Compounds and compositions for inhbiting the activity of shp2

ABSTRACT

The present invention relates to compounds of formula I: 
     
       
         
         
             
             
         
       
         
         
           
             in which Y 1 , Y 2 , Y 3 , Y 4  R 1 , R 2 , R 4a , R 4b , R 5a , R 5b , R 6a  and R 6b  are defined in the Summary of the Invention; capable of inhibiting the activity of SHP2. The invention further provides a process for the preparation of compounds of the invention, pharmaceutical preparations comprising such compounds and methods of using such compounds and compositions in the management of diseases or disorders associated with the aberrant activity of SHP2.

BACKGROUND Field of the Invention

The present invention relates to compounds capable of inhibiting theactivity of SHP2. The invention further provides a process for thepreparation of compounds of the invention, pharmaceutical preparationscomprising such compounds and methods of using such compounds andcompositions in the management of diseases or disorders associated withthe aberrant activity of SHP2.

Background of the Invention

The Src Homolgy-2 phosphatase (SHP2) is a non-receptor protein tyrosinephosphatase encoded by the PTPN11 gene that contributes to multiplecellular functions including proliferation, differentiation, cell cyclemaintenance and migration. SHP2 is involved in signaling through theRas-mitogen-activated protein kinase, the JAKSTAT or the phosphoinositol3-kinase-AKT pathways.

SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), acatalytic domain (PTP), and a C-terminal tail. The two SH2 domainscontrol the subcellular localization and functional regulation of SHP2.The molecule exists in an inactive, self-inhibited conformationstabilized by a binding network involving residues from both the N-SH2and PTP domains. Stimulation by, for example, cytokines or growthfactors leads to exposure of the catalytic site resulting in enzymaticactivation of SHP2.

Mutations in the PTPN11 gene and subsequently in SHP2 have beenidentified in several human diseases, such as Noonan Syndrome, LeopardSyndrome, juvenile myelomonocytic leukemias, neuroblastoma, melanoma,acute myeloid leukemia and cancers of the breast, lung and colon. SHP2,therefore, represents a highly attractive target for the development ofnovel therapies for the treatment of various diseases. The compounds ofthe present invention fulfill the need of small molecules to thatinhibit the activity of SHP2.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides compounds of Formula I:

in which: Y₁ is selected from N, NH and CH; Y₂ is selected from N, NHand CH; Y₃ is selected from N and C; Y₄ is selected from N and CH; R₁ isselected from R_(1a) and SR_(1a); wherein R_(1a) is selected fromphenyl, pyridinyl, pyrazine, pyridazine, 2,3-dihydrobenzofuran andpyrimidine; wherein said phenyl, pyridinyl, pyrazine, pyridazine orpyrimidine is unsubstituted or substituted with 1 to 3 R₄ groups;wherein each R₄ group is independently selected from halo, amino,—NH(C₁₋₄ alkyl), —N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl),morpholino, pyrrolidinyl, hydroxyl, C₁₋₃ alkyl, C₁₋₃ alkoxy andhalo-substituted-C₁₋₃alkyl; wherein said pyrrolidinyl of R₄ can beunsubstituted or substituted with hydroxy; R₂ is selected from H, CH₃and halo-substituted-C₁₋₂alkyl; R_(4a) and R_(4b) are each independentlyselected from hydrogen, hydroxy and fluoro; with proviso that R_(4a) andR_(4b) cannot both be OH; R_(5a) is selected from amino andamino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl, C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl, C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃ alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂ alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH.

In a second aspect, the present invention provides a pharmaceuticalcomposition which contains a compound of Formula I or a N-oxidederivative, tautomer, individual isomers and mixture of isomers thereof;or a pharmaceutically acceptable salt thereof, in admixture with one ormore suitable excipients.

In a third aspect, the present invention provides a method of treating adisease in an animal in which modulation of SHP2 activity can prevent,inhibit or ameliorate the pathology and/or symptomology of the diseases,which method comprises administering to the animal a therapeuticallyeffective amount of a compound of Formula I or a N-oxide derivative,individual isomers and mixture of isomers thereof, or a pharmaceuticallyacceptable salt thereof.

In a fourth aspect, the present invention provides a method of treatinga disease in an animal in which modulation of SHP2 activity can prevent,inhibit or ameliorate the pathology and/or symptomology of the diseases,which method comprises administering to the animal a therapeuticallyeffective amount of a compound of Formula I or a N-oxide derivative,individual isomers and mixture of isomers thereof, or a pharmaceuticallyacceptable salt thereof, in simultaneous or sequential combination withan anti-cancer therapeutic.

In a fifth aspect, the present invention provides the use of a compoundof Formula I in the manufacture of a medicament for treating a diseasein an animal in which SHP2 activity contributes to the pathology and/orsymptomology of the disease.

In a sixth aspect, the present invention provides a process forpreparing compounds of Formula I and the N-oxide derivatives, prodrugderivatives, protected derivatives, individual isomers and mixture ofisomers thereof, and the pharmaceutically acceptable salts thereof.

Definitions

The general terms used hereinbefore and hereinafter preferably havewithin the context of this disclosure the following meanings, unlessotherwise indicated, where more general terms wherever used may,independently of each other, be replaced by more specific definitions orremain, thus defining more detailed embodiments of the invention:

“Alkyl” refers to a fully saturated branched or unbranched hydrocarbonmoiety having up to 20 carbon atoms. Unless otherwise provided, alkylrefers to hydrocarbon moieties having 1 to 7 carbon atoms (C₁₋₇alkyl),or 1 to 4 carbon atoms (C₁₋₄alkyl). Representative examples of alkylinclude, but are not limited to, methyl, ethyl, n-propyl, iso-propyl,n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl,neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl,2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl and the like. Asubstituted alkyl is an alkyl group containing one or more, such as one,two or three substituents selected from halogen, hydroxy or alkoxygroups. Halo-substituted-alkyl and halo-substituted-alkoxy, can beeither straight-chained or branched and includes, methoxy, ethoxy,difluoromethyl, trifluoromethyl, pentafluoroethyl, difluoromethoxy,trifluoromethoxy, and the like.

“Aryl” means a monocyclic or fused bicyclic aromatic ring assemblycontaining six to ten ring carbon atoms. For example, aryl may be phenylor naphthyl, preferably phenyl. “Arylene” means a divalent radicalderived from an aryl group.

“Heteroaryl” is as defined for aryl above where one or more of the ringmembers is a heteroatom. For example C₅₋₁₀heteroaryl is a minimum of 5members as indicated by the carbon atoms but that these carbon atoms canbe replaced by a heteroatom. Consequently, C₅₋₁₀heteroaryl includespyridyl, indolyl, indazolyl, quinoxalinyl, quinolinyl, benzofuranyl,benzopyranyl, benzothiopyranyl, benzo[1,3]dioxole, imidazolyl,benzo-imidazolyl, pyrimidinyl, furanyl, oxazolyl, isoxazolyl, triazolyl,tetrazolyl, pyrazolyl, thienyl, etc.

“Cycloalkyl” means a saturated or partially unsaturated, monocyclic,fused bicyclic or bridged polycyclic ring assembly containing the numberof ring atoms indicated. For example, C₃₋₁₀cycloalkyl includescyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, etc.

“Heterocycloalkyl” means cycloalkyl, as defined in this application,provided that one or more of the ring carbons indicated, are replaced bya moiety selected from —O—, —NR—, —C(O)—, —S—, —S(O)— or —S(O)₂—,wherein R is hydrogen, C₁₋₄ alkyl or a nitrogen protecting group. Forexample, C₃₋₈heterocycloalkyl as used in this application to describecompounds of the invention includes morpholino, pyrrolidinyl,pyrrolidinyl-2-one, piperazinyl, piperidinyl, piperidinylone,1,4-dioxa-8-aza-spiro[4.5]dec-8-yl, thiomorpholino, sulfanomorpholino,sulfonomorpholino, etc.

“Halogen” (or halo) preferably represents chloro or fluoro, but may alsobe bromo or iodo.

“SHP2” means “Src Homolgy-2 phosphatase” and is also known as SH-PTP2,SH-PTP3, Syp, PTP1D, PTP2C, SAP-2 or PTPN11.

Cancers harboring “PTPN11 mutations” include but are not limited to:N58Y; D61Y, V; E69K; A72V, T, D; E76G, Q, K (ALL); G60A; D61Y; E69V;F71K; A72V; T73I; E76G, K; R289G; G503V (AML); G60R, D61Y, V, N; Y62D;E69K; A72T, V; T73I; E76K, V, G, A, Q; E139D; G503A, R; Q506P (JMML);G60V; D61V; E69K; F71L; A72V; E76A (MDS); Y63C (CMML); Y62C; E69K; T507K(neuroblastoma); V46L; N58S; E76V (Lung cancer); R138Q (melanoma); E76G(colon cancer).

Compounds of formula I may have different isomeric forms. For example,any asymmetric carbon atom may be present in the (R)-, (S)- or(R,S)-configuration, preferably in the (R)- or (S)-configuration.Substituents at a double bond or especially a ring may be present incis-(=Z-) or trans (=E-) form. The compounds may thus be present asmixtures of isomers or preferably as pure isomers, preferably as purediastereomers or pure enantiomers.

Where the plural form (e.g. compounds, salts) is used, this includes thesingular (e.g. a single compound, a single salt). “A compound” does notexclude that (e.g. in a pharmaceutical formulation) more than onecompound of the formula I (or a salt thereof) is present, the “a” merelyrepresenting the indefinite article. “A” can thus preferably be read as“one or more”, less preferably alternatively as “one”.

Wherever a compound or compounds of the formula I are mentioned, this isfurther also intended to include N-oxides of such compounds and/ortautomers thereof.

The term “and/or an N-oxide thereof, a tautomer thereof and/or a(preferably pharmaceutically acceptable) salt thereof” especially meansthat a compound of the formula I may be present as such or in mixturewith its N-oxide, as tautomer (e.g. due to keto-enol, lactam-lactim,amide-imidic acid or enamine-imine tautomerism) or in (e.g. equivalencyreaction caused) mixture with its tautomer, or as a salt of the compoundof the formula I and/or any of these forms or mixtures of two or more ofsuch forms.

The present invention also includes all suitable isotopic variations ofthe compounds of the invention, or pharmaceutically acceptable saltsthereof. An isotopic variation of a compound of the invention or apharmaceutically acceptable salt thereof is defined as one in which atleast one atom is replaced by an atom having the same atomic number butan atomic mass different from the atomic mass usually found in nature.Examples of isotopes that may be incorporated into the compounds of theinvention and pharmaceutically acceptable salts thereof include, but arenot limited to, isotopes of hydrogen, carbon, nitrogen and oxygen suchas ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ³⁵S, ¹⁸F, ³⁶Cl and ¹²³I.Certain isotopic variations of the compounds of the invention andpharmaceutically acceptable salts thereof, for example, those in which aradioactive isotope such as ³H or ¹⁴C is incorporated, are useful indrug and/or substrate tissue distribution studies. In particularexamples, ³H and ¹⁴C isotopes may be used for their ease of preparationand detectability. In other examples, substitution with isotopes such as²H may afford certain therapeutic advantages resulting from greatermetabolic stability, such as increased in vivo half-life or reduceddosage requirements. Isotopic variations of the compounds of theinvention or pharmaceutically acceptable salts thereof can generally beprepared by conventional procedures using appropriate isotopicvariations of suitable reagents.

DESCRIPTION OF PREFERRED EMBODIMENTS

The present invention relates to compounds capable of inhibiting theactivity of SHP2. In one aspect of the invention, with respect tocompounds of formula I, are compounds of Formulae Ia, Ib, Ic, Id, Ie, Ifand Ig:

in which: R₁ is selected from phenyl, 2,3-dihydrobenzofuran orpyridinyl; wherein said phenyl or pyridinyl is unsubstituted orsubstituted with 1 to 3 R₄ groups; wherein each R₄ group isindependently selected from halo, amino, —NH(C₁₋₄ alkyl), —N(C₁₋₄alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅ cycloalkyl), morpholino, pyrrolidinyl, C₁₋₃alkyl, C₁₋₃ alkoxy and halo-substituted-C₁₋₃ alkyl; wherein saidpyrrolidinyl of R₄ can be unsubstituted or substituted with hydroxy; R₂is selected from H and CH₃; R_(4a) and R_(4b) are each independentlyselected from hydrogen, hydroxy and fluoro; with proviso that R_(4a) andR_(4b) cannot both be OH; R_(5a) is selected from amino andamino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆alkyl or C₁₋₂alkoxy-substituted C₁₋₃alkyl of R_(5b) is unsubstitutedor substituted with 1-3 fluorines; or R_(5a) and R_(5b), together withthe carbon atom to which R_(5a) and R_(5b) are attached, form a groupselected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂alkyl andhydroxy-methyl; Rig is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; or thepharmaceutically acceptable salts thereof.

In another aspect of the invention are compounds of formula Ia:

in which: R₁ is phenyl substituted with 1 to 3 R₄ groups; wherein eachR₄ group is independently selected from halo, amino, —NH(C₁₋₄alkyl),—N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl), morpholino,pyrrolidinyl, C₁₋₃alkyl, C₁₋₃alkoxy and halo-substituted-C₁₋₃ alkyl;wherein said pyrrolidinyl of R₄ can be unsubstituted or substituted withhydroxy; R₂ is selected from H and CH₃; R_(4a) and R_(4b) are eachindependently selected from hydrogen, hydroxy and fluoro; with provisothat R_(4a) and R_(4b) cannot both be OH; R_(5a) is selected from aminoand amino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein said6alkyl or C₁₋₂ alkoxy-substituted C₁₋₃ alkyl of R_(5b) is unsubstitutedor substituted with 1-3 fluorines; or R_(5a) and R_(5b), together withthe carbon atom to which R_(5a) and R_(5b) are attached, form a groupselected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl, halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy andC₁₋₃alkoxy; and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; andR_(6a) and R_(6b) are each independently selected from hydrogen, hydroxyand fluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; orthe pharmaceutically acceptable salts thereof.

In a further aspect of the invention are compounds, or apharmaceutically acceptable salt thereof, selected from:

In another aspect of the invention are compounds of formula Ic:

in which: R₁ is selected from phenyl, 2,3-dihydrobenzofuran orpyridinyl; wherein said phenyl or pyridinyl is unsubstituted orsubstituted with 1 to 3 R₄ groups; wherein each R₄ group isindependently selected from halo, amino, —NH(C₁₋₄ alkyl), —N(C₁₋₄alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅ cycloalkyl), morpholino, pyrrolidinyl, C₁₋₃alkyl, C₁₋₃ alkoxy and halo-substituted-C₁₋₃alkyl; wherein saidpyrrolidinyl of R₄ can be unsubstituted or substituted with hydroxy; R₂is selected from H and CH₃; R_(4a) and R_(4b) are each independentlyselected from hydrogen, hydroxy and fluoro; with proviso that R_(4a) andR_(4b) cannot both be OH; R_(5a) is selected from amino andamino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃ alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂ alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; or thepharmaceutically acceptable salts thereof.

In a further aspect of the invention are compounds, or apharmaceutically acceptable salt thereof, selected from:

In another aspect of the invention are compounds of formula Id:

in which: R₁ is selected from phenyl, 2,3-dihydrobenzofuran orpyridinyl; wherein said phenyl or pyridinyl is unsubstituted orsubstituted with 1 to 3 R₄ groups; wherein each R₄ group isindependently selected from halo, amino, —NH(C₁₋₄alkyl), —N(C₁₋₄alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅ cycloalkyl), morpholino, pyrrolidinyl,C₁₋₃alkyl, C₁₋₃alkoxy and halo-substituted-C₁₋₃alkyl; wherein saidpyrrolidinyl of R₄ can be unsubstituted or substituted with hydroxy; R₂is selected from H and CH₃; R_(4a) and R_(4b) are each independentlyselected from hydrogen, hydroxy and fluoro; with proviso that R_(4a) andR_(4b) cannot both be OH; R_(5a) is selected from amino andamino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆alkyl or C₁₋₂alkoxy-substituted C₁₋₃alkyl of R_(5b) is unsubstitutedor substituted with 1-3 fluorines; or R_(5a) and R_(5b), together withthe carbon atom to which R_(5a) and R_(5b) are attached, form a groupselected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl, halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy andC₁₋₃alkoxy; and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; andR_(6a) and R_(6b) are each independently selected from hydrogen, hydroxyand fluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; orthe pharmaceutically acceptable salts thereof.

In a further aspect of the invention are compounds, or apharmaceutically acceptable salt thereof, selected from:

In a further aspect of the invention are compounds, or apharmaceutically acceptable salt thereof, selected from:

In another aspect of the invention are compounds, or pharmaceuticallyacceptable salts thereof, of formula II:

in which: Y₅ is selected from N and CH; Y₆ is selected from CH₂ and NH;R₁ is selected from R_(1a) and SR_(1a); wherein R_(1a) is selected fromphenyl, pyridinyl, pyrazine, pyridazine, 2,3-dihydrobenzofuran andpyrimidine; wherein said phenyl, pyridinyl, pyrazine, pyridazine orpyrimidine is unsubstituted or substituted with 1 to 3 R₄ groups;wherein each R₄ group is independently selected from halo, amino,—NH(C₁₋₄alkyl), —N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl),morpholino, pyrrolidinyl, hydroxyl, C₁₋₃alkyl, C₁₋₃alkoxy andhalo-substituted-C₁₋₃alkyl; wherein said pyrrolidinyl of R₄ can beunsubstituted or substituted with hydroxy; R_(4a) and R_(4b) are eachindependently selected from hydrogen, hydroxy and fluoro; with provisothat R_(4a) and R_(4b) cannot both be OH; R_(5a) is selected from aminoand amino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which Rya and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂ alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH.

In a further aspect of the invention are compounds, or apharmaceutically acceptable salt thereof, selected from:

Pharmacology and Utility

The Src Homolgy-2 phosphatase (SHP2) is a protein tyrosine phosphataseencoded by the PTPN11 gene that contributes to multiple cellularfunctions including proliferation, differentiation, cell cyclemaintenance and migration. SHP2 is involved in signaling through theRas-mitogen-activated protein kinase, the JAKSTAT or the phosphoinositol3-kinase-AKT pathways. SHP2 mediates activation of Erk1 and Erk2(Erk1/2, Erk) MAP kinases by receptor tyrosine kinases such as ErbB1,ErbB2 and c-Met.

SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), acatalytic domain (PTP), and a C-terminal tail. The two SH2 domainscontrol the subcellular localization and functional regulation of SHP2.The molecule exists in an inactive conformation, inhibiting its ownactivity via a binding network involving residues from both the N-SH2and PTP domains. In response to growth factor stimulation, SHP2 binds tospecific tyrosine-phosphorylated sites on docking proteins such as Gab1and Gab2 via its SH2 domains. This induces a conformational change thatresults in SHP2 activation.

Mutations in PTPN11 have been identified in several human diseases, suchas Noonan Syndrome, Leopard Syndrome, juvenile myelomonocytic leukemias,neuroblastoma, melanoma, acute myeloid leukemia and cancers of thebreast, lung and colon. SHP2 is an important downstream signalingmolecule for a variety of receptor tyrosine kinases, including thereceptors of platelet-derived growth factor (PDGF-R), fibroblast growthfactor (FGF-R) and epidermal growth factor (EGF-R). SHP2 is also animportant downstream signaling molecule for the activation of themitogen activated protein (MAP) kinase pathway which can lead to celltransformation, a prerequisite for the development of cancer. Knock-downof SHP2 significantly inhibited cell growth of lung cancer cell lineswith SHP2 mutation or EML4/ALK translocations as well as EGFR amplifiedbreast cancers and esophageal cancers. SHP2 is also activated downstreamof oncogenes in gastric carcinoma, anaplastic large-cell lymphoma andglioblastoma.

Noonan Syndrome (NS) and Leopard Syndrome (LS)—PTPN11 mutations cause LS(multiple lentigenes, electrocardiographic conduction abnormalities,ocular hypertelorism, pulmonic stenosis, abnormal genitalia, retardationof growth, sensorineural deafness) and NS (congenital anomaliesincluding cardiac defects, craniofacial abnormalities and shortstature). Both disorders are part of a family of autosomal dominantsyndromes caused by germline mutations in components of theRAS/RAF/MEK/ERK mitogen activating protein kinase pathway, required fornormal cell growth and differentiation. Aberrant regulation of thispathway has profound effects, particularly on cardiac development,resulting in various abnormalities, including valvuloseptal defectsand/or hypertrophic cardiomyopathy (HCM). Perturbations of the MAPKsignaling pathway have been established as central to these disordersand several candidate genes along this pathway have been identified inhumans, including mutations in KRAS, NRAS, SOS1, RAFT, BRAF, MEK1, MEK2,SHOC2, and CBL. The gene most commonly mutated in NS and LS is PTPN11.Germline mutations in PTPN11 (SHP2) are found in ˜50% of the cases withNS and nearly all patients with LS that shares certain features with NS.For NS, Y62D and Y63C substitutions in the protein are largely invariantand are among the most common mutations. Both these mutations affect thecatalytically inactive conformation of SHP2 without perturbing thebinding of the phosphatase to its phosphorylated signaling partners.

Juvenile Myelomonocytic Leukemias (JMML)—Somatic mutations in PTPN11(SHP2) occur in about 35% of the patients with JMML, a childhoodmyeloproliferative disorder (MPD). These gain-of-function mutations aretypically point mutations in the N-SH2 domain or in the phosphatasedomain, which prevent self-inhibition between the catalytic domain andthe N-SH2 domain, resulting in SHP2 activity.

Acute Myeloid Leukemia PTPN11 mutations have been identified in: ˜10% ofpediatric acute leukemias, such as myelodysplastic syndrome (MDS); ˜7%of B cell acute lymphoblastic leukemia (B-ALL); and ˜4% of acute myeloidleukemia (AML).

NS and leukemia mutations cause changes in amino acids located at theinterface formed by the N-SH2 and PTP domains in the self-inhibited SHP2conformation, disrupting the inhibitory intramolecular interaction,leading to hyperactivity of the catalytic domain.

SHP2 acts as a positive regulator in receptor tyrosine kinase (RTK)signaling. Cancers containing RTK alterations (EGFR^(amp), Her2^(amp),FGFR^(amp), Met^(amp), translocated/activated RTK, i.e. ALK, BCR/ABL)include Esophageal, Breast, Lung, Colon, Gastric, Glioma, Head and Neckcancers.

Esophageal cancer (or oesophageal cancer) is a malignancy of theesophagus. There are various subtypes, primarily squamous cell cancer(<50%) and adenocarcinoma. There is a high rate of RTK expression inesophageal adenocarcinoma and squamous cell cancer. A SHP2 inhibitor ofthe invention can, therefore, be employed for innovative treatmentstrategies.

Breast cancer is a major type of cancer and a leading cause of death inwomen, where patients develop resistance to current drugs. There arefour major subtypes of breast cancers including luminal A, luminal B,Her2 like, and triple negative/Basal-like. Triple negative breast cancer(TNBC) is an aggressive breast cancer lacking specific targeted therapy.Epidermal growth factor receptor I (EGFR) has emerged as a promisingtarget in TNBC. Inhibition of Her2 as well as EGFR via SHP2 may be apromising therapy in breast cancer.

Lung Cancer—NSCLC is currently a major cause of cancer-relatedmortality. accounting for about 85% of lung cancers (predominantlyadenocarcinomas and squamous cell carcinomas). Although cytotoxicchemotherapy remains an important part of treatment, targeted therapiesbased on genetic alterations such as EGFR and ALK in the tumor are morelikely to benefit from a targeted therapy.

Colon Cancer—Approximately 30% to 50% of colorectal tumors are known tohave a mutated (abnormal) KRAS, and BRAF mutations occur in 10 to 15% ofcolorectal cancers. For a subset of patients whose colorectal tumorshave been demonstrated to over express EGFR, these patients exhibit afavorable clinical response to anti-EGFR therapy.

Gastic Cancer is one of the most prevalent cancer types. Aberrantexpression of tyrosine kinases, as reflected by the aberrant tyrosinephosphorylation in gastric cancer cells, is known in the art. Threereceptor-tyrosine kinases, c-met (HGF receptor), FGF receptor 2, anderbB2/neu are frequently amplified in gastric carcinomas. Thus,subversion of different signal pathways may contribute to theprogression of different types of gastric cancers.

Neuroblastoma is a pediatric tumor of the developing sympathetic nervoussystem, accounting for about 8% of childhood cancers. Genomicalterations of the anaplastic lymphoma kinase (ALK) gene have beenpostulated to contribute to neuroblastoma pathogenesis.

Squamous-cell carcinoma of the head and neck (SCCHN). High levels ofEGFR expression are correlated with poor prognosis and resistance toradiation therapy in a variety of cancers, mostly in squamous-cellcarcinoma of the head and neck (SCCHN). Blocking of the EGFR signalingresults in inhibition of the stimulation of the receptor, cellproliferation, and reduced invasiveness and metastases. The EGFR is,therefore, a prime target for new anticancer therapy in SCCHN.

The present invention relates to compounds capable of inhibiting theactivity of SHP2. The invention further provides a process for thepreparation of compounds of the invention and pharmaceuticalpreparations comprising such compounds. Another aspect of the presentinvention relates to a method of treating SHP2-mediated disorderscomprising the step of administering to a patient in need thereof atherapeutically effective amount of a compound of formula I as definedin the Summary of the Invention.

In certain embodiments, the present invention relates to theaforementioned method, wherein said SHP2-mediated disorders are cancersselected from, but not limited to: JMML; AML; MDS; B-ALL; neuroblastoma;esophageal; breast cancer; lung cancer; colon cancer; Gastric cancer,Head and Neck cancer.

The compounds of the present invention may also be useful in thetreatment of other diseases or conditions related to the aberrantactivity of SHP2. Thus, as a further aspect, the invention relates to amethod of treatment of a disorder selected from: NS; LS; JMML; AML; MDS;B-ALL; neuroblastoma; esophageal; breast cancer; lung cancer; coloncancer; gastric cancer; head and neck cancer.

A SHP2 inhibitor of the present invention may be usefully combined withanother pharmacologically active compound, or with two or more otherpharmacologically active compounds, particularly in the treatment ofcancer. For example, a compound of the formula (I), or apharmaceutically acceptable salt thereof, as defined above, may beadministered simultaneously, sequentially or separately in combinationwith one or more agents selected from chemotherapy agents, for example,mitotic inhibitors such as a taxane, a vinca alkaloid, paclitaxel,docetaxel, vincristine, vinblastine, vinorelbine or vinflunine, andother anticancer agents, e.g. cisplatin, 5-fluorouracil or5-fluoro-2-4(1H,3H)-pyrimidinedione (5FU), flutamide or gemcitabine.

Such combinations may offer significant advantages, includingsynergistic activity, in therapy.

In certain embodiments, the present invention relates to theaforementioned method, wherein said compound is administeredparenterally.

In certain embodiments, the present invention relates to theaforementioned method, wherein said compound is administeredintramuscularly, intravenously, subcutaneously, orally, pulmonary,intrathecally, topically or intranasally.

In certain embodiments, the present invention relates to theaforementioned method, wherein said compound is administeredsystemically.

In certain embodiments, the present invention relates to theaforementioned method, wherein said patient is a mammal.

In certain embodiments, the present invention relates to theaforementioned method, wherein said patient is a primate.

In certain embodiments, the present invention relates to theaforementioned method, wherein said patient is a human.

In another aspect, the present invention relates to a method of treatingan SHP2-mediated disorder, comprising the step of: administering to apatient in need thereof a therapeutically effective amount of achemothereutic agent in combination with a therapeutically effectiveamount of a compound of formula I as defined in the Summary of theInvention.

Pharmaceutical Compositions

In another aspect, the present invention provides pharmaceuticallyacceptable compositions which comprise a therapeutically-effectiveamount of one or more of the compounds described above, formulatedtogether with one or more pharmaceutically acceptable carriers(additives) and/or diluents. As described in detail below, thepharmaceutical compositions of the present invention may be speciallyformulated for administration in solid or liquid form, including thoseadapted for the following: (1) oral administration, for example,drenches (aqueous or non-aqueous solutions or suspensions), tablets,e.g., those targeted for buccal, sublingual, and systemic absorption,boluses, powders, granules, pastes for application to the tongue; (2)parenteral administration, for example, by subcutaneous, intramuscular,intravenous or epidural injection as, for example, a sterile solution orsuspension, or sustained-release formulation; (3) topical application,for example, as a cream, ointment, or a controlled-release patch orspray applied to the skin; (4) intravaginally or intrarectally, forexample, as a pessary, cream or foam; (5) sublingually; (6) ocularly;(7) transdermally; (8) nasally; (9) pulmonary; or (10) intrathecally.

The phrase “therapeutically-effective amount” as used herein means thatamount of a compound, material, or composition comprising a compound ofthe present invention which is effective for producing some desiredtherapeutic effect in at least a sub-population of cells in an animal ata reasonable benefit/risk ratio applicable to any medical treatment.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically-acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or steric acid), orsolvent encapsulating material, involved in carrying or transporting thesubject compound from one organ, or portion of the body, to anotherorgan, or portion of the body. Each carrier must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand not injurious to the patient. Some examples of materials which canserve as pharmaceutically-acceptable carriers include: (1) sugars, suchas lactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) pH buffered solutions; (21)polyesters, polycarbonates and/or polyanhydrides; and (22) othernon-toxic compatible substances employed in pharmaceutical formulations.

As set out above, certain embodiments of the present compounds maycontain a basic functional group, such as amino or alkylamino, and are,thus, capable of forming pharmaceutically-acceptable salts withpharmaceutically-acceptable acids. The term “pharmaceutically-acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ in the administration vehicle or thedosage form manufacturing process, or by separately reacting a purifiedcompound of the invention in its free base form with a suitable organicor inorganic acid, and isolating the salt thus formed during subsequentpurification. Representative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate,valerate, oleate, palmitate, stearate, laurate, benzoate, lactate,phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate,napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonatesalts and the like. (See, for example, Berge et al. (1977)“Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19).

The pharmaceutically acceptable salts of the subject compounds includethe conventional nontoxic salts or quaternary ammonium salts of thecompounds, e.g., from non-toxic organic or inorganic acids. For example,such conventional nontoxic salts include those derived from inorganicacids such as hydrochloride, hydrobromic, sulfuric, sulfamic,phosphoric, nitric, and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicyclic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isothionic, and the like.

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically-acceptable salts with pharmaceutically-acceptablebases. The term “pharmaceutically-acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present invention. These salts can likewise beprepared in situ in the administration vehicle or the dosage formmanufacturing process, or by separately reacting the purified compoundin its free acid form with a suitable base, such as the hydroxide,carbonate or bicarbonate of a pharmaceutically-acceptable metal cation,with ammonia, or with a pharmaceutically-acceptable organic primary,secondary or tertiary amine. Representative alkali or alkaline earthsalts include the lithium, sodium, potassium, calcium, magnesium, andaluminum salts and the like. Representative organic amines useful forthe formation of base addition salts include ethylamine, diethylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.(See, for example, Berge et al., supra)

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically-acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral,nasal, topical (including buccal and sublingual), rectal, vaginal and/orparenteral administration. The formulations may conveniently bepresented in unit dosage form and may be prepared by any methods wellknown in the art of pharmacy. The amount of active ingredient which canbe combined with a carrier material to produce a single dosage form willvary depending upon the host being treated, the particular mode ofadministration. The amount of active ingredient which can be combinedwith a carrier material to produce a single dosage form will generallybe that amount of the compound which produces a therapeutic effect.Generally, out of one hundred percent, this amount will range from about0.1 percent to about ninety-nine percent of active ingredient,preferably from about 5 percent to about 70 percent, most preferablyfrom about 10 percent to about 30 percent.

In certain embodiments, a formulation of the present invention comprisesan excipient selected from the group consisting of cyclodextrins,celluloses, liposomes, micelle forming agents, e.g., bile acids, andpolymeric carriers, e.g., polyesters and polyanhydrides; and a compoundof the present invention. In certain embodiments, an aforementionedformulation renders orally bioavailable a compound of the presentinvention.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules, trouches and thelike), the active ingredient is mixed with one or morepharmaceutically-acceptable carriers, such as sodium citrate ordicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,certain silicates, and sodium carbonate; (5) solution retarding agents,such as paraffin; (6) absorption accelerators, such as quaternaryammonium compounds and surfactants, such as poloxamer and sodium laurylsulfate; (7) wetting agents, such as, for example, cetyl alcohol,glycerol monostearate, and non-ionic surfactants; (8) absorbents, suchas kaolin and bentonite clay; (9) lubricants, such as talc, calciumstearate, magnesium stearate, solid polyethylene glycols, sodium laurylsulfate, zinc stearate, sodium stearate, stearic acid, and mixturesthereof; (10) coloring agents; and (11) controlled release agents suchas crospovidone or ethyl cellulose. In the case of capsules, tablets andpills, the pharmaceutical compositions may also comprise bufferingagents. Solid compositions of a similar type may also be employed asfillers in soft and hard-shelled gelatin capsules using such excipientsas lactose or milk sugars, as well as high molecular weight polyethyleneglycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be formulated for rapid release,e.g., freeze-dried. They may be sterilized by, for example, filtrationthrough a bacteria-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedin sterile water, or some other sterile injectable medium immediatelybefore use. These compositions may also optionally contain opacifyingagents and may be of a composition that they release the activeingredient(s) only, or preferentially, in a certain portion of thegastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions which can be used include polymeric substancesand waxes. The active ingredient can also be in micro-encapsulated form,if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such carriers as are known in theart to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically-acceptablecarrier, and with any preservatives, buffers, or propellants which maybe required.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyimide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the compoundin a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsugars, alcohols, antioxidants, buffers, bacteriostats, solutes whichrender the formulation isotonic with the blood of the intended recipientor suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms upon the subject compounds may be ensuredby the inclusion of various antibacterial and antifungal agents, forexample, paraben, chlorobutanol, phenol sorbic acid, and the like. Itmay also be desirable to include isotonic agents, such as sugars, sodiumchloride, and the like into the compositions. In addition, prolongedabsorption of the injectable pharmaceutical form may be brought about bythe inclusion of agents which delay absorption such as aluminummonostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally-administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissue.

When the compounds of the present invention are administered aspharmaceuticals, to humans and animals, they can be given per se or as apharmaceutical composition containing, for example, 0.1 to 99% (morepreferably, 10 to 30%) of active ingredient in combination with apharmaceutically acceptable carrier.

The preparations of the present invention may be given orally,parenterally, topically, or rectally. They are of course given in formssuitable for each administration route. For example, they areadministered in tablets or capsule form, by injection, inhalation, eyelotion, ointment, suppository, etc. administration by injection,infusion or inhalation; topical by lotion or ointment; and rectal bysuppositories. Oral administrations are preferred.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

These compounds may be administered to humans and other animals fortherapy by any suitable route of administration, including orally,nasally, as by, for example, a spray, rectally, intravaginally,parenterally, intracisternally and topically, as by powders, ointmentsor drops, including buccally and sublingually.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically-acceptable dosage forms by conventional methodsknown to those of skill in the art.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient which is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion ormetabolism of the particular compound being employed, the rate andextent of absorption, the duration of the treatment, other drugs,compounds and/or materials used in combination with the particularcompound employed, the age, sex, weight, condition, general health andprior medical history of the patient being treated, and like factorswell known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound which is the lowest dose effective toproduce a therapeutic effect. Such an effective dose will generallydepend upon the factors described above. Generally, oral, intravenous,intracerebroventricular and subcutaneous doses of the compounds of thisinvention for a patient, when used for the indicated analgesic effects,will range from about 0.0001 to about 100 mg per kilogram of body weightper day.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms. Preferred dosing is one administrationper day.

While it is possible for a compound of the present invention to beadministered alone, it is preferable to administer the compound as apharmaceutical formulation (composition).

The compounds according to the invention may be formulated foradministration in any convenient way for use in human or veterinarymedicine, by analogy with other pharmaceuticals.

In another aspect, the present invention provides pharmaceuticallyacceptable compositions which comprise a therapeutically-effectiveamount of one or more of the subject compounds, as described above,formulated together with one or more pharmaceutically acceptablecarriers (additives) and/or diluents. As described in detail below, thepharmaceutical compositions of the present invention may be speciallyformulated for administration in solid or liquid form, including thoseadapted for the following: (1) oral administration, for example,drenches (aqueous or non-aqueous solutions or suspensions), tablets,boluses, powders, granules, pastes for application to the tongue; (2)parenteral administration, for example, by subcutaneous, intramuscularor intravenous injection as, for example, a sterile solution orsuspension; (3) topical application, for example, as a cream, ointmentor spray applied to the skin, lungs, or mucous membranes; or (4)intravaginally or intrarectally, for example, as a pessary, cream orfoam; (5) sublingually or buccally; (6) ocularly; (7) transdermally; or(8) nasally.

The term “treatment” is intended to encompass also prophylaxis, therapyand cure.

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

The compound of the invention can be administered as such or inadmixtures with pharmaceutically acceptable carriers and can also beadministered in conjunction with antimicrobial agents such aspenicillins, cephalosporins, aminoglycosides and glycopeptides.Conjunctive therapy, thus includes sequential, simultaneous and separateadministration of the active compound in a way that the therapeuticaleffects of the first administered one is not entirely disappeared whenthe subsequent is administered.

Microemulsification technology can improve bioavailability of somelipophilic (water insoluble) pharmaceutical agents. Examples includeTrimetrine (Dordunoo, S. K., et al., Drug Development and IndustrialPharmacy, 17(12), 1685-1713, 1991 and REV 5901 (Sheen, P. C., et al., JPharm Sci 80(7), 712-714, 1991). Among other things, microemulsificationprovides enhanced bioavailability by preferentially directing absorptionto the lymphatic system instead of the circulatory system, which therebybypasses the liver, and prevents destruction of the compounds in thehepatobiliary circulation.

While all suitable amphiphilic carriers are contemplated, the presentlypreferred carriers are generally those that haveGenerally-Recognized-as-Safe (GRAS) status, and that can both solubilizethe compound of the present invention and microemulsify it at a laterstage when the solution comes into a contact with a complex water phase(such as one found in human gastrointestinal tract). Usually,amphiphilic ingredients that satisfy these requirements have HLB(hydrophilic to lipophilic balance) values of 2-20, and their structurescontain straight chain aliphatic radicals in the range of C-6 to C-20.Examples are polyethylene-glycolized fatty glycerides and polyethyleneglycols.

Commercially available amphiphilic carriers are particularlycontemplated, including Gelucire-series, Labrafil, Labrasol, orLauroglycol (all manufactured and distributed by Gattefosse Corporation,Saint Priest, France), PEG-mono-oleate, PEG-di-oleate, PEG-mono-laurateand di-laurate, Lecithin, Polysorbate 80, etc (produced and distributedby a number of companies in USA and worldwide).

Hydrophilic polymers suitable for use in the present invention are thosewhich are readily water-soluble, can be covalently attached to avesicle-forming lipid, and which are tolerated in vivo without toxiceffects (i.e., are biocompatible). Suitable polymers includepolyethylene glycol (PEG), polylactic (also termed polylactide),polyglycolic acid (also termed polyglycolide), a polylactic-polyglycolicacid copolymer, and polyvinyl alcohol. Preferred polymers are thosehaving a molecular weight of from about 100 or 120 daltons up to about5,000 or 10,000 daltons, and more preferably from about 300 daltons toabout 5,000 daltons. In a particularly preferred embodiment, the polymeris polyethyleneglycol having a molecular weight of from about 100 toabout 5,000 daltons, and more preferably having a molecular weight offrom about 300 to about 5,000 daltons. In a particularly preferredembodiment, the polymer is polyethyleneglycol of 750 daltons (PEG(750)).Polymers may also be defined by the number of monomers therein; apreferred embodiment of the present invention utilizes polymers of atleast about three monomers, such PEG polymers consisting of threemonomers (approximately 150 daltons).

Other hydrophilic polymers which may be suitable for use in the presentinvention include polyvinylpyrrolidone, polymethoxazoline,polyethyloxazoline, polyhydroxypropyl methacrylamide,polymethacrylamide, polydimethylacrylamide, and derivatized cellulosessuch as hydroxymethylcellulose or hydroxyethylcellulose.

In certain embodiments, a formulation of the present invention comprisesa biocompatible polymer selected from the group consisting ofpolyamides, polycarbonates, polyalkylenes, polymers of acrylic andmethacrylic esters, polyvinyl polymers, polyglycolides, polysiloxanes,polyurethanes and co-polymers thereof, celluloses, polypropylene,polyethylenes, polystyrene, polymers of lactic acid and glycolic acid,polyanhydrides, poly(ortho)esters, poly(butic acid), poly(valeric acid),poly(lactide-co-caprolactone), polysaccharides, proteins, polyhyaluronicacids, polycyanoacrylates, and blends, mixtures, or copolymers thereof.

Cyclodextrins are cyclic oligosaccharides, consisting of 6, 7 or 8glucose units, designated by the Greek letter alpha, beta or gamma,respectively. Cyclodextrins with fewer than six glucose units are notknown to exist. The glucose units are linked by alpha-1,4-glucosidicbonds. As a consequence of the chair conformation of the sugar units,all secondary hydroxyl groups (at C-2, C-3) are located on one side ofthe ring, while all the primary hydroxyl groups at C-6 are situated onthe other side. As a result, the external faces are hydrophilic, makingthe cyclodextrins water-soluble. In contrast, the cavities of thecyclodextrins are hydrophobic, since they are lined by the hydrogen ofatoms C-3 and C-5, and by ether-like oxygens. These matrices allowcomplexation with a variety of relatively hydrophobic compounds,including, for instance, steroid compounds such as 17.beta.-estradiol(see, e.g., van Uden et al. Plant Cell Tiss. Org. Cult. 38:1-3-113(1994)). The complexation takes place by Van der Waals interactions andby hydrogen bond formation. For a general review of the chemistry ofcyclodextrins, see, Wenz, Agnew. Chem. Int. Ed. Engl., 33:803-822(1994).

The physico-chemical properties of the cyclodextrin derivatives dependstrongly on the kind and the degree of substitution. For example, theirsolubility in water ranges from insoluble (e.g.,triacetyl-beta-cyclodextrin) to 147% soluble (w/v)(G-2-beta-cyclodextrin). In addition, they are soluble in many organicsolvents. The properties of the cyclodextrins enable the control oversolubility of various formulation components by increasing or decreasingtheir solubility.

Numerous cyclodextrins and methods for their preparation have beendescribed. For example, Parmeter (I), et al. (U.S. Pat. No. 3,453,259)and Gramera, et al. (U.S. Pat. No. 3,459,731) described electroneutralcyclodextrins. Other derivatives include cyclodextrins with cationicproperties [Parmeter (II), U.S. Pat. No. 3,453,257], insolublecrosslinked cyclodextrins (Solms, U.S. Pat. No. 3,420,788), andcyclodextrins with anionic properties [Parmeter (III), U.S. Pat. No.3,426,011]. Among the cyclodextrin derivatives with anionic properties,carboxylic acids, phosphorous acids, phosphinous acids, phosphonicacids, phosphoric acids, thiophosphonic acids, thiosulphinic acids, andsulfonic acids have been appended to the parent cyclodextrin [see,Parmeter (III), supra]. Furthermore, sulfoalkyl ether cyclodextrinderivatives have been described by Stella, et al. (U.S. Pat. No.5,134,127).

Liposomes consist of at least one lipid bilayer membrane enclosing anaqueous internal compartment. Liposomes may be characterized by membranetype and by size Small unilamellar vesicles (SUVs) have a singlemembrane and typically range between 0.02 and 0.05 μm in diameter; largeunilamellar vesicles (LUVS) are typically larger than 0.05 μmOligolamellar large vesicles and multilamellar vesicles have multiple,usually concentric, membrane layers and are typically larger than 0.1μm. Liposomes with several nonconcentric membranes, i.e., severalsmaller vesicles contained within a larger vesicle, are termedmultivesicular vesicles.

One aspect of the present invention relates to formulations comprisingliposomes containing a compound of the present invention, where theliposome membrane is formulated to provide a liposome with increasedcarrying capacity. Alternatively or in addition, the compound of thepresent invention may be contained within, or adsorbed onto, theliposome bilayer of the liposome. The compound of the present inventionmay be aggregated with a lipid surfactant and carried within theliposome's internal space; in these cases, the liposome membrane isformulated to resist the disruptive effects of the activeagent-surfactant aggregate.

According to one embodiment of the present invention, the lipid bilayerof a liposome contains lipids derivatized with polyethylene glycol(PEG), such that the PEG chains extend from the inner surface of thelipid bilayer into the interior space encapsulated by the liposome, andextend from the exterior of the lipid bilayer into the surroundingenvironment.

Active agents contained within liposomes of the present invention are insolubilized form. Aggregates of surfactant and active agent (such asemulsions or micelles containing the active agent of interest) may beentrapped within the interior space of liposomes according to thepresent invention. A surfactant acts to disperse and solubilize theactive agent, and may be selected from any suitable aliphatic,cycloaliphatic or aromatic surfactant, including but not limited tobiocompatible lysophosphatidylcholines (LPCs) of varying chain lengths(for example, from about C.sub.14 to about C.sub.20).Polymer-derivatized lipids such as PEG-lipids may also be utilized formicelle formation as they will act to inhibit micelle/membrane fusion,and as the addition of a polymer to surfactant molecules decreases theCMC of the surfactant and aids in micelle formation. Preferred aresurfactants with CMCs in the micromolar range; higher CMC surfactantsmay be utilized to prepare micelles entrapped within liposomes of thepresent invention, however, micelle surfactant monomers could affectliposome bilayer stability and would be a factor in designing a liposomeof a desired stability.

Liposomes according to the present invention may be prepared by any of avariety of techniques that are known in the art. See, e.g., U.S. Pat.No. 4,235,871; Published PCT applications WO 96/14057; New RRC,Liposomes: A practical approach, IRL Press, Oxford (1990), pages 33-104;Lasic DD, Liposomes from physics to applications, Elsevier SciencePublishers BV, Amsterdam, 1993.

For example, liposomes of the present invention may be prepared bydiffusing a lipid derivatized with a hydrophilic polymer into preformedliposomes, such as by exposing preformed liposomes to micelles composedof lipid-grafted polymers, at lipid concentrations corresponding to thefinal mole percent of derivatized lipid which is desired in theliposome. Liposomes containing a hydrophilic polymer can also be formedby homogenization, lipid-field hydration, or extrusion techniques, asare known in the art.

In one aspect of the present invention, the liposomes are prepared tohave substantially homogeneous sizes in a selected size range. Oneeffective sizing method involves extruding an aqueous suspension of theliposomes through a series of polycarbonate membranes having a selecteduniform pore size; the pore size of the membrane will correspond roughlywith the largest sizes of liposomes produced by extrusion through thatmembrane. See e.g., U.S. Pat. No. 4,737,323 (Apr. 12, 1988).

The release characteristics of a formulation of the present inventiondepend on the encapsulating material, the concentration of encapsulateddrug, and the presence of release modifiers. For example, release can bemanipulated to be pH dependent, for example, using a pH sensitivecoating that releases only at a low pH, as in the stomach, or a higherpH, as in the intestine. An enteric coating can be used to preventrelease from occurring until after passage through the stomach. Multiplecoatings or mixtures of cyanamide encapsulated in different materialscan be used to obtain an initial release in the stomach, followed bylater release in the intestine. Release can also be manipulated byinclusion of salts or pore forming agents, which can increase wateruptake or release of drug by diffusion from the capsule. Excipientswhich modify the solubility of the drug can also be used to control therelease rate. Agents which enhance degradation of the matrix or releasefrom the matrix can also be incorporated. They can be added to the drug,added as a separate phase (i.e., as particulates), or can beco-dissolved in the polymer phase depending on the compound. In allcases the amount should be between 0.1 and thirty percent (w/w polymer).Types of degradation enhancers include inorganic salts such as ammoniumsulfate and ammonium chloride, organic acids such as citric acid,benzoic acid, and ascorbic acid, inorganic bases such as sodiumcarbonate, potassium carbonate, calcium carbonate, zinc carbonate, andzinc hydroxide, and organic bases such as protamine sulfate, spermine,choline, ethanolamine, diethanolamine, and triethanolamine andsurfactants such as Tween® and Pluronic®. Pore forming agents which addmicrostructure to the matrices (i.e., water soluble compounds such asinorganic salts and sugars) are added as particulates. The range shouldbe between one and thirty percent (w/w polymer).

Uptake can also be manipulated by altering residence time of theparticles in the gut. This can be achieved, for example, by coating theparticle with, or selecting as the encapsulating material, a mucosaladhesive polymer. Examples include most polymers with free carboxylgroups, such as chitosan, celluloses, and especially polyacrylates (asused herein, polyacrylates refers to polymers including acrylate groupsand modified acrylate groups such as cyanoacrylates and methacrylates).

Pharmaceutical Combinations

The invention especially relates to the use of a compound of the formulaI (or a pharmaceutical composition comprising a compound of the formulaI) in the treatment of one or more of the diseases mentioned herein;wherein the response to treatment is beneficial as demonstrated, forexample, by the partial or complete removal of one or more of thesymptoms of the disease up to complete cure or remission.

A compound of formula (I) can also be used in combination with thefollowing compounds and antibody-drug conjugates:

BCR-ABL inhibitors: Imatinib (Gleevec®); Inilotinib hydrochloride;Nilotinib (Tasigna®); Dasatinib (BMS-345825); Bosutinib (SKI-606);Ponatinib (AP24534); Bafetinib (INNO406); Danusertib (PHA-739358),AT9283 (CAS 1133385-83-7); Saracatinib (AZD0530); andN-[2-[(1S,4R)-6-[[4-(Cyclobutylamino)-5-(trifluoromethyl)-2-pyrimidinyl]amino]-1,2,3,4-tetrahydronaphthalen-1,4-imin-9-yl]-2-oxoethyl]-acetamide(PF-03814735, CAS 942487-16-3).

ALK inhibitors: PF-2341066 (XALKORI®; crizotinib);5-chloro-N4-(2-(isopropylsulfonyl)phenyl)-N2-(2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)pyrimidine-2,4-diamine;GSK1838705A; and CH5424802.

BRAF inhibitors: Vemurafanib (PLX4032); and Dabrafenib.

FLT3 inhibitors sunitinib malate (sold under the tradename Sutent® byPfizer); PKC412 (midostaurin); tanutinib, sorafenib, sunitinib,midostaurin, lestaurtinib, KW-2449, quizartinib (AC220) and crenolanib.

MEK Inhibitors trametinib.

Vascular Endothelial Growth Factor (VEGF) receptor inhibitors:Bevacizumab (sold under the trademark Avastin® by Genentech/Roche),axitinib,(N-methyl-2-[[3-[(E)-2-pyridin-2-ylethenyl]-1H-indazol-6-yl]sulfanyl]benzamide,also known as AG013736, and described in PCT Publication No. WO01/002369), Brivanib Alaninate((S)-((R)-1-(4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate,also known as BMS-582664), motesanib(N-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide,and described in PCT Publication No. WO 02/066470), pasireotide (alsoknown as SOM230, and described in PCT Publication No. WO 02/010192),sorafenib (sold under the tradename Nexavar®);

HER2 receptor inhibitors: Trastuzumab (sold under the trademarkHerceptin® by Genentech/Roche), neratinib (also known as HKI-272,(2E)-N-[4-[[3-chloro-4-[(pyridin-2-yl)methoxy]phenyl]amino]-3-cyano-7-ethoxyquinolin-6-yl]-4-(dimethylamino)but-2-enamide,and described PCT Publication No. WO 05/028443), lapatinib or lapatinibditosylate (sold under the trademark Tykerb® by GlaxoSmithKline);Trastuzumab emtansine (in the United States, ado-trastuzumab emtansine,trade name Kadcyla)—an antibody-drug conjugate consisting of themonoclonal antibody trastuzumab (Herceptin) linked to the cytotoxicagent mertansine (DM1);

CD20 antibodies: Rituximab (sold under the trademarks Riuxan® andMabThera® by Genentech/Roche), tositumomab (sold under the trademarksBexxar® by GlaxoSmithKline), ofatumumab (sold under the trademarkArzerra® by GlaxoSmithKline);

Tyrosine kinase inhibitors: Erlotinib hydrochloride (sold under thetrademark Tarceva® by Genentech/Roche), Linifanib(N-[4-(3-amino-1H-indazol-4-yl)phenyl]-N′-(2-fluoro-5-methylphenyl)urea,also known as ABT 869, available from Genentech), sunitinib malate (soldunder the tradename Sutent® by Pfizer), bosutinib(4-[(2,4-dichloro-5-methoxyphenyl)amino]-6-methoxy-7-[3-(4-methylpiperazin-1-yl)propoxy]quinoline-3-carbonitrile,also known as SKI-606, and described in U.S. Pat. No. 6,780,996),dasatinib (sold under the tradename Sprycel® by Bristol-Myers Squibb),armala (also known as pazopanib, sold under the tradename Votrient® byGlaxoSmithKline), imatinib and imatinib mesylate (sold under thetradenames Gilvec® and Gleevec® by Novartis);

DNA Synthesis inhibitors: Capecitabine (sold under the trademark Xeloda®by Roche), gemcitabine hydrochloride (sold under the trademark Gemzar®by Eli Lilly and Company), nelarabine((2R,3S,4R,5R)-2-(2-amino-6-methoxy-purin-9-yl)-5-(hydroxymethyl)oxolane-3,4-diol,sold under the tradenames Arranon® and Atriance® by GlaxoSmithKline);

Antineoplastic agents: oxaliplatin (sold under the tradename Eloxatin®ay Sanofi-Aventis and described in U.S. Pat. No. 4,169,846);

Epidermal growth factor receptor (EGFR) inhibitors: Gefitnib (sold underthe tradename Iressa®),N-4-[4-[(3-Chloro-4-fluorophenyl)amino]-7-[[(3″S″)-tetrahydro-3-furanyl]oxy]-6-quinazolinyl]-4(dimethylamino)-2-butenamide,sold under the tradename Tovok® by Boehringer Ingelheim), cetuximab(sold under the tradename Erbitux® by Bristol-Myers Squibb), panitumumab(sold under the tradename Vectibix® by Amgen);

HER dimerization inhibitors: Pertuzumab (sold under the trademarkOmnitarg@, by Genentech);

Human Granulocyte colony-stimulating factor (G-CSF) modulators:Filgrastim (sold under the tradename Neupogen® by Amgen);

Immunomodulators: Afutuzumab (available from Roche®), pegfilgrastim(sold under the tradename Neulasta® by Amgen), lenalidomide (also knownas CC-5013, sold under the tradename Revlimid®), thalidomide (sold underthe tradename Thalomid®);

CD40 inhibitors: Dacetuzumab (also known as SGN-40 or huS2C6, availablefrom Seattle Genetics, Inc);

Pro-apoptotic receptor agonists (PARAS): Dulanermin (also known asAMG-951, available from Amgen/Genentech);

Hedgehog antagonists:2-chloro-N-[4-chloro-3-(2-pyridinyl)phenyl]-4-(methylsulfonyl)-benzamide(also known as GDC-0449, and described in PCT Publication No. WO06/028958);

PI3K inhibitors:4-[2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine(also known as GDC 0941 and described in PCT Publication Nos. WO09/036082 and WO 09/055730),2-Methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile(also known as BEZ 235 or NVP-BEZ 235, and described in PCT PublicationNo. WO 06/122806);

Phospholipase A2 inhibitors: Anagrelide (sold under the tradenameAgrylin®);

BCL-2 inhibitors:4-[4-[[2-(4-chlorophenyl)-5,5-dimethyl-1-cyclohexen-1-yl]methyl]-1-piperazinyl]-N-[[4-[[(1R)-3-(4-morpholinyl)-1-[(phenylthio)methyl]propyl]amino]-3-[(trifluoromethyl)sulfonyl]phenyl]sulfonyl]benzamide(also known as ABT-263 and described in PCT Publication No. WO09/155386);

Mitogen-activated protein kinase kinase (MEK) inhibitors: XL-518 (CasNo. 1029872-29-4, available from ACC Corp.);

Aromatase inhibitors: Exemestane (sold under the trademark Aromasin® byPfizer), letrozole (sold under the tradename Femara® by Novartis),anastrozole (sold under the tradename Arimidex®);

Topoisomerase I inhibitors: Irinotecan (sold under the trademarkCamptosar® by Pfizer), topotecan hydrochloride (sold under the tradenameHycamtin® by GlaxoSmithKline);

Topoisomerase II inhibitors: etoposide (also known as VP-16 andEtoposide phosphate, sold under the tradenames Toposar®, VePesid® andEtopophos®), teniposide (also known as VM-26, sold under the tradenameVumon®);

mTOR inhibitors: Temsirolimus (sold under the tradename Torisel® byPfizer), ridaforolimus (formally known as deferolimus,(1R,2R,4S)-4-[(2R)-2[1R,9S,12S,15R,16E,18R,19R,21R,23S,24E,26E,28Z,30S,32S,35R)-1,18-dihydroxy-19,30-dimethoxy-15,17,21,23,29,35-hexamethyl-2,3,10,14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30.3.1.0^(4,9)]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]-2-methoxycyclohexyldimethylphosphinate, also known as AP23573 and MK8669, and described inPCT Publication No. WO 03/064383), everolimus (sold under the tradenameAfinitor® by Novartis);

Osteoclastic bone resorption inhibitors:1-Hydroxy-2-imidazol-1-yl-phosphonoethyl) phosphonic acid monohydrate(sold under the tradename Zometa® by Novartis);

CD33 Antibody Drug Conjugates: Gemtuzumab ozogamicin (sold under thetradename Mylotarg® by Pfizer/Wyeth);

CD22 Antibody Drug Conjugates: Inotuzumab ozogamicin (also referred toas CMC-544 and WAY-207294, available from Hangzhou Sage Chemical Co.,Ltd.);

CD20 Antibody Drug Conjugates: Ibritumomab tiuxetan (sold under thetradename Zevalin®);

Somatostain analogs: octreotide (also known as octreotide acetate, soldunder the tradenames Sandostatin® and Sandostatin LAW));

Synthetic Interleukin-11 (IL-11): oprelvekin (sold under the tradenameNeumega® by Pfizer/Wyeth);

Synthetic erythropoietin: Darbepoetin alfa (sold under the tradenameAranesp® by Amgen);

Receptor Activator for Nuclear Factor κ B (RANK) inhibitors: Denosumab(sold under the tradename Prolia® by Amgen);

Thrombopoietin mimetic peptibodies: Romiplostim (sold under thetradename Nplate® by Amgen;

Cell growth stimulators: Palifermin (sold under the tradename Kepivance®by Amgen);

Anti-Insulin-like Growth Factor-1 receptor (IGF-1R) antibodies:Figitumumab (also known as CP-751,871, available from ACC Corp),robatumumab (CAS No. 934235-44-6);

Anti-CS1 antibodies: Elotuzumab (HuLuc63, CAS No. 915296-00-3);

CD52 antibodies: Alemtuzumab (sold under the tradename Campath®);

CTLA-4 inhibitors: Tremelimumab (IgG2 monoclonal antibody available fromPfizer, formerly known as ticilimumab, CP-675,206), ipilimumab (CTLA-4antibody, also known as MDX-010, CAS No. 477202-00-9);

Histone deacetylase inhibitors (HDI): Voninostat (sold under thetradename Zolinza® by Merck);

Alkylating agents: Temozolomide (sold under the tradenames Temodar® andTemodal® by Schering-Plough/Merck), dactinomycin (also known asactinomycin-D and sold under the tradename Cosmegen®), melphalan (alsoknown as L-PAM, L-sarcolysin, and phenylalanine mustard, sold under thetradename Alkeran®), altretamine (also known as hexamethylmelamine(HMM), sold under the tradename Hexalen®), carmustine (sold under thetradename BiCNU®), bendamustine (sold under the tradename Treanda®),busulfan (sold under the tradenames Busulfex® and Myleran®), carboplatin(sold under the tradename Paraplatin®), lomustine (also known as CCNU,sold under the tradename CeeNU®), cisplatin (also known as CDDP, soldunder the tradenames Platinol® and Platinol®-AQ), chlorambucil (soldunder the tradename Leukeran®), cyclophosphamide (sold under thetradenames Cytoxan® and Neosar®), dacarbazine (also known as DTIC, DICand imidazole carboxamide, sold under the tradename DTIC-Dome®),altretamine (also known as hexamethylmelamine (HMM) sold under thetradename Hexalen®), ifosfamide (sold under the tradename Ifex®),procarbazine (sold under the tradename Matulane®), mechlorethamine (alsoknown as nitrogen mustard, mustine and mechloroethamine hydrochloride,sold under the tradename Mustargen®), streptozocin (sold under thetradename Zanosar®), thiotepa (also known as thiophosphoamide, TESPA andTSPA, sold under the tradename Thioplex®;

Biologic response modifiers: bacillus calmette-guerin (sold under thetradenames theraCys® and TICE® BCG), denileukin diftitox (sold under thetradename Ontak®);

Anti-tumor antibiotics: doxorubicin (sold under the tradenamesAdriamycin® and Rubex®), bleomycin (sold under the tradename Lenoxane®),daunorubicin (also known as dauorubicin hydrochloride, daunomycin, andrubidomycin hydrochloride, sold under the tradename Cerubidine®),daunorubicin liposomal (daunorubicin citrate liposome, sold under thetradename DaunoXome®), mitoxantrone (also known as DHAD, sold under thetradename Novantrone®), epirubicin (sold under the tradename Ellence™),idarubicin (sold under the tradenames Idamycin®, Idamycin PFS®),mitomycin C (sold under the tradename Mutamycin®);

Anti-microtubule agents: Estramustine (sold under the tradename Emcyl®);

Cathepsin K inhibitors: Odanacatib (also know as MK-0822,N-(1-cyanocyclopropyl)-4-fluoro-N²-{(1S)-2,2,2-trifluoro-1-[4′-(methylsulfonyl)biphenyl-4-yl]ethyl}-L-leucinamide,available from Lanzhou Chon Chemicals, ACC Corp., and ChemieTek, anddescribed in PCT Publication no. WO 03/075836);

Epothilone B analogs: Ixabepilone (sold under the tradename Lxempra® byBristol-Myers Squibb);

Heat Shock Protein (HSP) inhibitors: Tanespimycin(17-allylamino-17-demethoxygeldanamycin, also known as KOS-953 and17-AAG, available from SIGMA, and described in U.S. Pat. No. 4,261,989);

TpoR agonists: Eltrombopag (sold under the tradenames Promacta® andRevolade® by GlaxoSmithKline);

Anti-mitotic agents: Docetaxel (sold under the tradename Taxotere® bySanofi-Aventis);

Adrenal steroid inhibitors: aminoglutethimide (sold under the tradenameCytadren®);

Anti-androgens: Nilutamide (sold under the tradenames Nilandron® andAnandron®), bicalutamide (sold under tradename Casodex®), flutamide(sold under the tradename Fulexin™);

Androgens: Fluoxymesterone (sold under the tradename Halotestin®);

Proteasome inhibitors: Bortezomib (sold under the tradename Velcade®);

CDK1 inhibitors: Alvocidib (also known as flovopirdol or HMR-1275,2-(2-chlorophenyl)-5,7-dihydroxy-8-[3S,4R)-3-hydroxy-1-methyl-4-piperidinyl]-4-chromenone,and described in U.S. Pat. No. 5,621,002);

Gonadotropin-releasing hormone (GnRH) receptor agonists: Leuprolide orleuprolide acetate (sold under the tradenames Viadure® by Bayer AG,Eligard® by Sanofi-Aventis and Lupron® by Abbott Lab);

Taxane anti-neoplastic agents: Cabazitaxel(1-hydroxy-7β,10β-dimethoxy-9-oxo-5β,20-epoxytax-11-ene-2α,4,13α-triyl-4-acetate-2-benzoate-13-[(2R,3S)-3-{[tert-butoxy)carbonyl]amino}-2-hydroxy-3-phenylpropanoate),larotaxel((2α,3ξ,4α,5β,7α,10β,13α)-4,10-bis(acetyloxy)-13-({(2R,3S)-3-[(tert-butoxycarbonyl)amino]-2-hydroxy-3-phenylpropanoyl}oxy)-1-hydroxy-9-oxo-5,20-epoxy-7,19-cyclotax-11-en-2-ylbenzoate);

5HT1a receptor agonists: Xaliproden (also known as SR57746,142-[(2-naphthyl)ethyl]-4[3-(trifluoromethyl)phenyl]-1,2,3,6-tetrahydropyridine,and described in U.S. Pat. No. 5,266,573);

HPC vaccines: Cervarix® sold by GlaxoSmithKline, Gardasil® sold byMerck;

Iron Chelating agents: Deferasinox (sold under the tradename Exjade® byNovartis);

Anti-metabolites: Claribine (2-chlorodeoxyadenosine, sold under thetradename Leustatin®), 5-fluorouracil (sold under the tradenameAdrucil®), 6-thioguanine (sold under the tradename Purinethol®),pemetrexed (sold under the tradename Alimta®), cytarabine (also known asarabinosylcytosine (Ara-C), sold under the tradename Cytosar-U®),cytarabine liposomal (also known as Liposomal Ara-C, sold under thetradename DepoCyt™), decitabine (sold under the tradename Dacogen®),hydroxyurea (sold under the tradenames Hydrea®, Droxia™ and Mylocel™),fludarabine (sold under the tradename Fludara®), floxuridine (sold underthe tradename FUDR®), cladribine (also known as 2-chlorodeoxyadenosine(2-CdA) sold under the tradename Leustatin™), methotrexate (also knownas amethopterin, methotrexate sodium (MTX), sold under the tradenamesRheumatrex® and Trexall™), pentostatin (sold under the tradenameNipent®);

Bisphosphonates: Pamidronate (sold under the tradename Aredia®),zoledronic acid (sold under the tradename Zometa®);

Demethylating agents: 5-azacitidine (sold under the tradename Vidaza®),decitabine (sold under the tradename Dacogen®);

Plant Alkaloids: Paclitaxel protein-bound (sold under the tradenameAbraxane®), vinblastine (also known as vinblastine sulfate,vincaleukoblastine and VLB, sold under the tradenames Alkaban-AQ® andVelban®), vincristine (also known as vincristine sulfate, LCR, and VCR,sold under the tradenames Oncovin® and Vincasar Pfs®), vinorelbine (soldunder the tradename Navelbine®), paclitaxel (sold under the tradenamesTaxol and Onxal™);

Retinoids: Alitretinoin (sold under the tradename Panretin®), tretinoin(all-trans retinoic acid, also known as ATRA, sold under the tradenameVesanoid®), Isotretinoin (13-cis-retinoic acid, sold under thetradenames Accutane®, Amnesteem®, Claravis®, Glarus®, Decutan®,Isotane®, Izotech®, Oratane®, Isotret®, and Sotret®), bexarotene (soldunder the tradename Targretin®);

Glucocorticosteroids: Hydrocortisone (also known as cortisone,hydrocortisone sodium succinate, hydrocortisone sodium phosphate, andsold under the tradenames Ala-Cort®, Hydrocortisone Phosphate,Solu-Cortef®, Hydrocort Acetate® and Lanacort®), dexamethazone((8S,9R,10S,11S,13S,14S,16R,17R)-9-fluoro-11,17-dihydroxy-17-(2-hydroxyacetyl)-10,13,16-trimethyl-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-3-one),prednisolone (sold under the tradenames Delta-Cortel®, Orapred®,Pediapred® and Prelone®), prednisone (sold under the tradenamesDeltasone®, Liquid Red®, Meticorten® and Orasone®), methylprednisolone(also known as 6-Methylprednisolone, Methylprednisolone Acetate,Methylprednisolone Sodium Succinate, sold under the tradenamesDuralone®, Medralone®, Medrol®, M-Prednisol® and Solu-Medrol®);

Cytokines: interleukin-2 (also known as aldesleukin and IL-2, sold underthe tradename Proleukin®), interleukin-11 (also known as oprevelkin,sold under the tradename Neumega®), alpha interferon alfa (also known asIFN-alpha, sold under the tradenames Intron® A, and Roferon-A®);

Estrogen receptor downregulators: Fulvestrant (sold under the tradenameFaslodex®);

Anti-estrogens: tamoxifen (sold under the tradename Novaldex®);

Toremifene (sold under the tradename Fareston®);

Selective estrogen receptor modulators (SERMs): Raloxifene (sold underthe tradename Evista®);

Leutinizing hormone releasing hormone (LHRH) agonists: Goserelin (soldunder the tradename Zoladex®);

Progesterones: megestrol (also known as megestrol acetate, sold underthe tradename Megace®);

Miscellaneous cytotoxic agents: Arsenic trioxide (sold under thetradename Trisenox®), asparaginase (also known as L-asparaginase,Erwinia L-asparaginase, sold under the tradenames Elspar® andKidrolase®);

A compound of formula (I) can also be used in combination with thefollowing adjunct therapies:

Anti-nausea drugs: NK-1 receptor antagonists: Casopitant (sold under thetradenames Rezonic® and Zunrisa® by GlaxoSmithKline); and

Cytoprotective agents: Amifostine (sold under the tradename Ethyol®),leucovorin (also known as calcium leucovorin, citrovorum factor andfolinic acid).

None of the quotations of references made within the present disclosureis to be understood as an admission that the references cited are priorart that would negatively affect the patentability of the presentinvention.

Processes for Making Compounds of the Invention

The present invention also includes processes for the preparation ofcompounds of the invention. In the reactions described, it can benecessary to protect reactive functional groups, for example hydroxy,amino, imino, thio or carboxy groups, where these are desired in thefinal product, to avoid their unwanted participation in the reactions.Conventional protecting groups can be used in accordance with standardpractice, for example, see T. W. Greene and P. G. M. Wuts in “ProtectiveGroups in Organic Chemistry”, John Wiley and Sons, 1991.

Compounds of Formula I can be prepared by proceeding as in the followingReaction Scheme I:

in which X, Y₁, Y₂, Y₃, Y₄, R₁, R₂, R_(4a), R_(4b), R_(5a), R_(5b),R_(6a) and R_(6b) are as defined by the Summary of the Invention.Compounds of formula I can be prepared by reacting a compound of formula2 with a compound of formula 3 in the presence of a suitable solvent(such as MeCN, DMF, or the like), a suitable coupling agent (such asBOP-Cl, BOP, or the like) and a suitable catalyst (such as DBU, or thelike). The reaction proceeds at a temperature range of about 15° C. toabout 100° C. and can take from about 1 hour to about 24 hours tocomplete.

in which X, Y₁, Y₂, Y₃, R₁, R₂, R_(4a), R_(4b), R_(5a), R_(5b), R_(6a)and R_(6b) are as defined by the Summary of the Invention and Q is aleaving group such as methyl-sulfonyl, chloro, or the like. Compounds offormula I can be prepared by reacting a compound of formula 4 with acompound of formula 3 in the presence of a suitable solvent (such asDMSO, NMP, or the like) and a suitable base (such as DIPEA, or thelike). The reaction proceeds at a temperature range of about 80° C. toabout 140° C. and can take from about 1 hour to about 24 hours tocomplete.

Compounds of Formula I, in which Y₃ is carbon, can be prepared byproceeding as in the following Reaction Scheme III:

in which X, Y₁, Y₂, Y₄, R₁, R₂, R_(4a), R_(4b), R_(5a), R_(5b), R_(6a)and R_(6b) are as defined by the Summary of the Invention. Compounds offormula I can be prepared by reacting a compound of formula 5 with acompound of formula 6 in the presence of a suitable solvent (such asTHF, or the like), a suitable catalyst (such asChloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II),or the like) and a suitable buffering agent (such as potassiumphosphate, or the like). The reaction proceeds at a temperature range ofabout 15° C. to about 100° C. and can take from about 1 hour to about 24hours to complete.

Detailed examples of the synthesis of compounds of Formula I can befound in the Examples, infra.

Additional Processes for Making Compounds of the Invention

A compound of the invention can be prepared as a pharmaceuticallyacceptable acid addition salt by reacting the free base form of thecompound with a pharmaceutically acceptable inorganic or organic acid.Alternatively, a pharmaceutically acceptable base addition salt of acompound of the invention can be prepared by reacting the free acid formof the compound with a pharmaceutically acceptable inorganic or organicbase.

Compounds of the formula I can also be modified by appending appropriatefunctionalities to enhance selective biological properties.Modifications of this kind are known in the art and include those thatincrease penetration into a given biological system (e.g. blood,lymphatic system, central nervous system, testis), increasebioavailability, increase solubility to allow parenteral administration(e.g. injection, infusion), alter metabolism and/or alter the rate ofsecretion. Examples of this type of modifications include but are notlimited to esterification, e.g. with polyethylene glycols,derivatisation with pivaloyloxy or fatty acid substituents, conversionto carbamates, hydroxylation of aromatic rings and heteroatomsubstitution in aromatic rings. Wherever compounds of the formula I,and/or N-oxides, tautomers and/or (preferably pharmaceuticallyacceptable) salts thereof are mentioned, this comprises such modifiedformulae, while preferably the molecules of the formula I, theirN-oxides, their tautomers and/or their salts are meant.

Alternatively, the salt forms of the compounds of the invention can beprepared using salts of the starting materials or intermediates. In viewof the close relationship between the novel compounds of the formula Iin free form and those in the form of their salts, including those saltsthat can be used as intermediates, for example in the purification oridentification of the novel compounds, any reference to the compounds ora compound of the formula I hereinbefore and hereinafter is to beunderstood as referring to the compound in free form and/or also to oneor more salts thereof, as appropriate and expedient, as well as to oneor more solvates, e.g. hydrates.

Salts are formed, for example, as acid addition salts, preferably withorganic or inorganic acids, from compounds of formula I with a basicnitrogen atom, especially the pharmaceutically acceptable salts.Suitable inorganic acids are, for example, halogen acids, such ashydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organicacids are, for example, carboxylic, phosphonic, sulfonic or sulfamicacids, for example acetic acid, propionic acid, octanoic acid, decanoicacid, dodecanoic acid, glycolic acid, lactic acid, fumaric acid,succinic acid, malonic acid, adipic acid, pimelic acid, suberic acid,azelaic acid, malic acid, tartaric acid, citric acid, amino acids, suchas glutamic acid or aspartic acid, maleic acid, hydroxymaleic acid,methylmaleic acid, cyclohexanecarboxylic acid, adamantanecarboxylicacid, benzoic acid, salicylic acid, 4-aminosalicylic acid, phthalicacid, phenylacetic acid, mandelic acid, cinnamic acid, methane- orethane-sulfonic acid, 2-hydroxyethanesulfonic acid,ethane-1,2-disulfonic acid, benzenesulfonic acid, 4-toluenesulfonicacid, 2-naphthalenesulfonic acid, 1,5-naphthalene-disulfonic acid, 2- or3-methylbenzenesulfonic acid, methylsulfuric acid, ethylsulfuric acid,dodecylsulfuric acid, N-cyclohexylsulfamic acid, N-methyl-, N-ethyl- orN-propyl-sulfamic acid, or other organic protonic acids, such asascorbic acid.

For isolation or purification purposes it is also possible to usepharmaceutically unacceptable salts, for example picrates orperchlorates. For therapeutic use, only pharmaceutically acceptablesalts or free compounds are employed (where applicable in the form ofpharmaceutical preparations), and these are therefore preferred.

The free acid or free base forms of the compounds of the invention canbe prepared from the corresponding base addition salt or acid additionsalt from, respectively. For example a compound of the invention in anacid addition salt form can be converted to the corresponding free baseby treating with a suitable base (e g, ammonium hydroxide solution,sodium hydroxide, and the like). A compound of the invention in a baseaddition salt form can be converted to the corresponding free acid bytreating with a suitable acid (e.g., hydrochloric acid, etc.).

Compounds of the invention in unoxidized form can be prepared fromN-oxides of compounds of the invention by treating with a reducing agent(e.g., sulfur, sulfur dioxide, triphenyl phosphine, lithium borohydride,sodium borohydride, phosphorus trichloride, tribromide, or the like) ina suitable inert organic solvent (e.g. acetonitrile, ethanol, aqueousdioxane, or the like) at 0 to 80° C.

Prodrug derivatives of the compounds of the invention can be prepared bymethods known to those of ordinary skill in the art (e.g., for furtherdetails see Saulnier et al., (1994), Bioorganic and Medicinal ChemistryLetters, Vol. 4, p. 1985). For example, appropriate prodrugs can beprepared by reacting a non-derivatized compound of the invention with asuitable carbamylating agent (e.g., 1,1-acyloxyalkylcarbanochloridate,para-nitrophenyl carbonate, or the like).

Protected derivatives of the compounds of the invention can be made bymeans known to those of ordinary skill in the art. A detaileddescription of techniques applicable to the creation of protectinggroups and their removal can be found in T. W. Greene, “ProtectingGroups in Organic Chemistry”, 3^(rd) edition, John Wiley and Sons, Inc.,1999.

Compounds of the present invention can be conveniently prepared, orformed during the process of the invention, as solvates (e.g.,hydrates). Hydrates of compounds of the present invention can beconveniently prepared by recrystallization from an aqueous/organicsolvent mixture, using organic solvents such as dioxin, tetrahydrofuranor methanol.

Compounds of the invention can be prepared as their individualstereoisomers by reacting a racemic mixture of the compound with anoptically active resolving agent to form a pair of diastereoisomericcompounds, separating the diastereomers and recovering the opticallypure enantiomers. While resolution of enantiomers can be carried outusing covalent diastereomeric derivatives of the compounds of theinvention, dissociable complexes are preferred (e.g., crystallinediastereomeric salts). Diastereomers have distinct physical properties(e.g., melting points, boiling points, solubilities, reactivity, etc.)and can be readily separated by taking advantage of thesedissimilarities. The diastereomers can be separated by chromatography,or preferably, by separation/resolution techniques based upondifferences in solubility. The optically pure enantiomer is thenrecovered, along with the resolving agent, by any practical means thatwould not result in racemization. A more detailed description of thetechniques applicable to the resolution of stereoisomers of compoundsfrom their racemic mixture can be found in Jean Jacques, Andre Collet,Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John WileyAnd Sons, Inc., 1981.

In summary, the compounds of Formula I can be made by a process, whichinvolves:

(a) that of reaction schemes I, II and III; and

(b) optionally converting a compound of the invention into apharmaceutically acceptable salt;

(c) optionally converting a salt form of a compound of the invention toa non-salt form;

(d) optionally converting an unoxidized form of a compound of theinvention into a pharmaceutically acceptable N-oxide;

(e) optionally converting an N-oxide form of a compound of the inventionto its unoxidized form;

(f) optionally resolving an individual isomer of a compound of theinvention from a mixture of isomers;

(g) optionally converting a non-derivatized compound of the inventioninto a pharmaceutically acceptable prodrug derivative; and

(h) optionally converting a prodrug derivative of a compound of theinvention to its non-derivatized form.

Insofar as the production of the starting materials is not particularlydescribed, the compounds are known or can be prepared analogously tomethods known in the art or as disclosed in the Examples hereinafter.

One of skill in the art will appreciate that the above transformationsare only representative of methods for preparation of the compounds ofthe present invention, and that other well known methods can similarlybe used.

Examples

The following examples and intermediates serve to illustrate theinvention without limiting the scope thereof. Some abbreviations used inthe examples are as follows: acetic acid (AcOH); acetonitrile (MeCN);triethylamine (TEA); tetrahydrofuran (THF); aqueous (aq); saturated(sat.); atmosphere (atm.);2,2′-bis-diphenylphosphanyl-[1,1′]binaphthalenyl (BINAP);4-dimethylaminopyridine (DMAP); tert-butoxycarbonyl (Boc);1,1-carbonyldiimidazole (CDI); di-tert-butyl dicarbonate (Boc₂O);benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphoniumhexafluorophosphate (BOP); dichloromethane (DCM); diethyl ether (Et₂O);p-toluene sulfonic acid (PTSA); ethyl acetate (EtOAc); ethanol (EtOH);lithium bis(trimethylsilyl)amide (LHMDS); diisopropyl azodicarboxylate(DIAD); N,N-diisopropyl-ethylamine (DIEA or DIPEA);N,N-dimethylformamide (DMF); dimethyl sulfoxide (DMSO);diphenylphosphoryl azide (DPPA); hour(s) (h);2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (HATU); High Performance Liquid Chromatography(HPLC); isopropyl alcohol (IPA); lithium aluminium hydride (LAH); liquidchromatography coupled with mass spectrometry (LCMS); lithiumdiisopropylamide (LDA); methanol (MeOH); milliliter(s) (mL); minute(s)(min); microwave (MW); sodium bis(trimethylsilyl)amide (NHMDS);n-butyllithium (n-BuLi);1,1-bis(diphenylphosphino)-ferrocenedichloropalladium (II)(PdCl₂(dppf)); tris(dibenzylideneacetone)dipalladium (0) (Pd₂(dba)₃);dichlorobis(triphenylphosphine)palladium (II) (PdCl₂(PPh3)₂); roomtemperature (RT); tetra-n-butylammonium fluoride (TBAF);tert-butyldimethylsilyl chloride (TBSCl); trifluoroacetic acid (TFA);tetrahydrofuran (THF); thin layer chromatography (TLC); retention time(t_(R)); (S)-(−)-2,2′-Bis(di-p-tolylphosphino)-1,1′-binaphthyl((S)-TolBINAP); & 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene(XantPhos).

Intermediate 1 (S)-7-azaspiro[3.5]nonan-1-amine

(a) A solution of tert-butyl 1-oxo-7-azaspiro[3.5]nonane-7-carboxylate(5.24 g, 21.9 mmol), titanium(IV) isopropoxide (16.2 mL, 54.7 mmol), and(R)-2-methylpropane-2-sulfinamide (3.45 g, 28.5 mmol) in THF (99 mL) wasstirred for 12 h at 65° C. After cooling to −78° C., MeOH (9.9 mL) wasadded followed by lithium borohydride (1.43 g, 65.7 mmol). The resultingmixture was stirred at −78° C. for 3 h and at room temperature for 1 h.MeOH was slowly added to quench the excess of borohydride followed byaddition of brine. The resulting mixture was stirred for 15 min and thenfiltered through Celite. The aqueous mixture was extracted with EtOAc(3×20 mL). The organic phases were dried over MgSO₄, filtered, andconcentrated. The resulting residue was purified by silicachromatography (0-50% EtOAc/heptane eluent) to provide (S)-tert-butyl1-((R)-1,1-dimethylethylsulfinamido)-7-azaspiro[3.5]nonane-7-carboxylate(4.79 g. 13.90 mmol) as a white solid. LCMS: m/z 345.3 (M+H)⁺.

(b) A solution of (S)-tert-butyl1-((R)-1,1-dimethylethylsulfinamido)-7-azaspiro[3.5]nonane-7-carboxylate(0.4 g, 1.16 mmol) in TFA (450 μL, 5.81 mmol) and DCM (3.5 mL) wasstirred for 30 min at 40° C. Sat. aq Na₂CO₃ was added until pH ˜11 andthe aqueous mixture was extracted with DCM (3×15 mL). The combinedorganic phases were washed with brine, dried over Na₂SO₄, filtered, andthe volatiles were removed under reduced pressure to give(R)-2-methyl-N-((S)-7-azaspiro[3.5]nonan-1-yl)propane-2-sulfinamide (237mg, 0.97 mmol) as a white solid. LCMS: n/z 245.5 (M+H)⁺.

(c) A solution of(R)-2-methyl-N-((S)-7-azaspiro[3.5]nonan-1-yl)propane-2-sulfinamide (200mg, 0.58 mmol) in DCM was treated with HCl (4.0 M in dioxane, 1.45 mL,5.81 mmol). The mixture was stirred at 40° C. for 1 h. At completion,the reaction was treated with sat. aq Na₂CO₃ and layers were separated.The aqueous phase was extracted with DCM. The combined organics werewashed with brine, dried over Na₂SO₄, filtered and concentrated toprovide (S)-7-azaspiro[3.5]nonan-1-amine (124 mg, 0.58 mmol, 100%yield).

Intermediate 2 (S)-7-azaspiro[3.5]nonan-1-amine

(a) A mixture of tert-butyl 4-formylpiperidine-1-carboxylate (35.0 g,164 mmol), lithium tert-butoxyde (15.77 g, 197 mmol), and allylbromide(11.54 mL, 189 mmol) in DMF (328 mL) was stirred for 1 h at 0° C. Themixture was poured into a separation funnel containing sat. aq NH₄Cl:H₂O(1:1, 500 mL) and it was extracted with Et₂O (5×50 mL). The combinedorganic phases were dried over MgSO₄, filtered, and the volatiles wereremoved under reduced pressure. The resulting residue was purified bysilica chromatography (0-25% EtOAc/heptane eluent) to give tert-butyl4-allyl-4-formylpiperidine-1-carboxylate (24 g, 95 mmol) as colorlessoil. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 9.52 (s, 1H), 5.53-5.76 (m,1H), 4.96-5.19 (m, 2H), 3.80 (br. s., 2H), 2.97 (t, J=11.49 Hz, 2H),2.26 (d, J=7.33 Hz, 2H), 1.95 (dt, J=13.71, 3.13 Hz, 2H), 1.38-1.58 (m,11H).

(b) To a solution of tert-butyl 4-allyl-4-formylpiperidine-1-carboxylate(24 g, 95 mmol) in THF (300 mL) was added (at −78° C. and under N₂)vinyl magnesium bromide (1 M in THF, 118 mL, 118 mmol). The resultingsolution was warmed to room temperature within 1 h. The mixture waspoured into a separation funnel containing sat. aq NH₄Cl (250 mL) andextracted with EtOAc (4×50 mL). The combined organic phases were driedover MgSO₄, filtered and the volatiles were removed under reducedpressure to give tert-butyl4-allyl-4-(1-hydroxyallyl)piperidine-1-carboxylate (26.7 g, 95 mmol) ascolorless oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 9.52 (s, 1H),5.56-5.75 (m, 1H), 5.05-5.18 (m, 2H), 3.80 (br. s., 2H), 2.97 (t,J=11.49 Hz, 2H), 2.26 (d, J=7.33 Hz, 2H), 1.96 (dt, J=13.83, 3.06 Hz,2H), 1.49-1.60 (m, 2H), 1.41-1.49 (m, 9H). This compound was used innext step without further purification.

(c) A mixture of tert-butyl4-allyl-4-(1-hydroxyallyl)piperidine-1-carboxylate (26.7 g, 95 mmol) andDess-Martin periodinane (44.3 g, 105 mmol) in DCM (380 mL) was stirredfor 1 h at room temperature. The mixture was poured into a separationfunnel containing sat. aq NaHCO₃:Na₂SO₃ (1:1, 300 mL) and it wasextracted with DCM (4×50 mL). The combined organic phases were driedover MgSO₄, filtered, and the volatiles were removed under reducedpressure to provide a white solid. This solid was suspended in heptane(250 mL) and sonicated for 5 min. The white suspension was filteredthrough a pad of Celite and the volatiles were removed under reducedpressure to give tert-butyl 4-acryloyl-4-allylpiperidine-1-carboxylate(26.5 g, 95 mmol) as yellow oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm6.81 (dd, J=16.93, 10.36 Hz, 1H), 6.40 (dd, J=16.80, 1.89 Hz, 1H), 5.71(dd, J=10.36, 2.02 Hz, 1H), 5.46-5.66 (m, 1H), 4.91-5.14 (m, 2H), 3.78(br. s., 2H), 2.96 (br. s., 2H), 2.25-2.39 (m, 2H), 1.97-2.15 (m, 2H),1.37-1.57 (m, 11H). This compound was used in next step without furtherpurification.

(d) To a solution of tert-butyl4-acryloyl-4-allylpiperidine-1-carboxylate (26.5 g, 95 mmol) in toluene(degassed, 850 mL) was added Grubbs II catalyst (2.02 g, 2.38 mmol) intoluene (degassed, 100 mL). The resulting mixture was stirred for 45 minat 85° C. The solvent was concentrated and the resulting residue waspurified by silica chromatography (0-40% EtOAc/heptane eluent) to givetert-butyl 1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate (20.76 g, 83mmol) as brown solid. A solution of this compound and2,3-dichloro-5,6-dicyanobenzoquinone (565 mg, 2.49 mmol) in toluene (540mL) was stirred for 15 min at room temperature. The resulting bright redsolution was filtered through a pad of Celite. Charcoal (200 g) wasadded and the resulting suspension was stirred for 2 h at roomtemperature. The mixture was filtered through a pad of Celite and thevolatiles were removed under reduce pressure. The resulting residue waspurified by silica chromatography (0-40% EtOAc/heptane eluent) to givetert-butyl 1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate (15.6 g, 62.3mmol) as white solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.63-7.74 (m,1 H), 6.20 (dt, J=5.81, 2.15 Hz, 1H), 3.99-4.25 (m, 2H), 2.92 (t,J=11.62 Hz, 2H), 2.63 (s, 2H), 1.72-1.86 (m, 2H), 1.49 (s, 9H), 1.29 (d,J=12.88 Hz, 2H).

Intermediate 3 (1R,3R)-3-methyl-8-azaspiro[4.5]decan-1-amine

(a) To a suspension of tert-butyl1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate (4.2 g, 16.71 mmol) and CuI(6.37 g, 33.4 mmol) in Et₂O (100 mL) was added (at 0° C. and under Na)MeLi (1.6 M in THF, 31.3 mL, 50.1 mmol). After stirring for 90 min at 0°C., the mixture was poured into a separation funnel containing sat. aqNH₄Cl and it was extracted with EtOAc (3×15 mL). The combined organicphases were dried over MgSO₄, filtered and the volatiles were removedunder reduced pressure. The resulting residue was purified by silicachromatography (0-50% EtOAc/heptane eluent) to give tert-butyl3-methyl-1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate (4.23 g, 15.82mmol) as colorless oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 3.89-4.00(m, 1H), 3.83 (d, J=13.39 Hz, 1H), 3.11 (ddd, J=13.64, 10.36, 3.28 Hz,1H), 2.99 (ddd, J=13.58, 10.42, 3.54 Hz, 1H), 2.47-2.59 (m, 1H),2.19-2.36 (m, 2H), 1.74-1.97 (m, 2H), 1.50-1.65 (m, 2H), 1.48 (s, 9H),1.33-1.44 (m, 2H), 1.17 (d, J=6.32 Hz, 3H).

(b) A mixture of tert-butyl3-methyl-1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate (4.23 g, 15.82mmol) and TFA (17 mL) in DCM (80 mL) was stirred for 30 min at roomtemperature. The volatiles were removed under reduced pressure. Amixture of the resulting residue, DIPEA (13.82 mL, 79 mmol), and benzylchloroformate (3.39 mL, 23.73 mmol) in DCM (80 mL) was stirred for 16 hat room temperature. The mixture was poured into a separation funnelcontaining sat. aq NH₄Cl and it was extracted with DCM (3×25 mL).

The combined organic phases were dried over MgSO₄, filtered and thevolatiles were removed under reduced pressure. The resulting residue waspurified by flash chromatography over silica gel (0-40% EtOAc/heptaneeluent) to give benzyl3-methyl-1-oxo-8-azaspiro[4.5]decane-8-carboxylate (4.58 g, 15.20 mmol)as light yellow oil. LCMS: m/z 302.2 (M+H)⁺.

(c) Benzyl 3-methyl-1-oxo-8-azaspiro[4.5]decane-8-carboxylate (4.58 g,15.20 mmol) was further purified by chiral SFC as follows: column: IA21×250 mm, flow rate: 70 g per minute, mobile phase: 45% (9:1 EtOH:MeCN)in CO₂, detection: 220 nm UV to give (R)-benzyl3-methyl-1-oxo-8-azaspiro[4.5]decane-8-carboxylate (2.02 g, 6.70 mmol)R_(t): 2.0 min; and (S)-benzyl3-methyl-1-oxo-8-azaspiro[4.5]decane-8-carboxylate (2.11 g, 7.0 mmol),R_(t)=3.6 min.

(d) A solution of (R)-benzyl3-methyl-1-oxo-8-azaspiro[4.5]decane-8-carboxylate (2.02 g, 6.70 mmol),titanium(IV) ethoxide (5.62 mL, 26.8 mmol), and(R)-2-methylpropane-2-sulfinamide (1.625 g, 13.4 mmol) in THF (67 mL)was stirred for 16 h at 65° C. After cooling to −78° C., MeOH (12 mL)was added followed by lithium borohydride (0.438 g, 20.11 mmol). Theresulting mixture was stirred for 16 h at −78° C. to room temperature.Sat. aq NH₄Cl was slowly added to quench the excess of borohydridefollowed by addition of EtOAc (100 mL). The resulting mixture wasvigorously stirred for 15 min and then filtered through a pad of Celite.The volatiles were removed under reduced pressure and the resultingresidue was purified by silica chromatography (5-90% EtOAc/heptaneeluent) to give (1R,3R)-benzyl1-((R)-1,1-dimethylethylsulfinamido)-3-methyl-8-azaspiro[4.5]decane-8-carboxylate(1.94 g, 4.77 mmol) as a white solid. LCMS: m/z 407.3 (M+H)⁺.

(e) A mixture of (1R,3R)-benzyl1-((R)-1,1-dimethylethylsulfinamido)-3-methyl-8-azaspiro[4.5]decane-8-carboxylate(100 mg, 0.246 mmol) and HCl (4 M in dioxane, 1.5 mL, 6.5 mmol) in MeOH(1.5 mL) was stirred in a microwave reactor for 14 h at 140° C. Aftercooling to room temperature, the volatiles were removed under reducedpressure to give (1R,3R)-3-methyl-8-azaspiro[4.5]decan-1-amine which wasused in next step without further purification. LCMS: m/z 169.2 (M+H)⁺.

Intermediate 4 (S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-2-oxa-8-azaspiro[4.5]decane-8-carboxylate

(a) A solution of tert-butyl4-hydroxy-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (544 mg, 2.11 mmol)and Dess-Martin periodinane (1.39 g, 3.17 mmol) in DCM (10 mL) wasstirred for 2 h at 0° C. At completion, 1:1 sat. aq NaHCO₃:sat. aqNa₂S₂O₃ (10 mL) was added. The mixture was stirred vigorously, then theorganic phase was separated. The aqueous phase was extracted with DCM(3×10 mL). The combined organic phases were dried over Na₂SO₄, filteredand concentrated. The resulting residue was purified by flashchromatography over silica gel (0-50% EtOAc/heptane eluent) to givetert-butyl 4-oxo-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (470 mg, 1.84mmol) as a colorless oil which crystallized upon standing. LCMS: m/z256.2 (M+H)⁺; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.08 (s, 2H), 4.05(s, 2H), 3.88 (dt, J=13.7, 4.9 Hz, 2H), 3.12 (ddd, J=13.6, 9.8, 3.6 Hz,2H), 1.75 (ddd, J=13.9, 9.7, 4.2 Hz, 2H), 1.58-1.51 (m, 2H), 1.48 (s,9H).

(b) A solution of tert-butyl4-oxo-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (220 mg, 0.86 mmol),titanium(IV) ethoxide (725 μL, 3.45 mmol), and(R)-2-methylpropane-2-sulfinamide (209 mg, 1.72 mmol) in THF (4 mL) wasstirred for 1 h at 90° C. After cooling to 0° C., lithium borohydride(23 mg, 1.06 mmol) was added. After stirring for 30 min, the reactionmixture was quenched by addition of MeOH. The volatiles were removedunder reduce pressure. The resulting residue was diluted with brine andextracted with EtOAc (4×10 mL). The combined organic phases were driedover Na₂SO₄, filtered and concentrated. The resulting residue waspurified by flash chromatography over silica gel (0-100% EtOAc/heptaneeluent) to give (S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-2-oxa-8-azaspiro[4.5]decane-8-carboxylate(170 mg, 0.47 mmol). LCMS: m/z 361.1 (M+H)⁺.

(c) A solution of (S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-2-oxa-8-azaspiro[4.5]decane-8-carboxylate(165 mg, 0.44 mmol) in MeOH was treated with HCl (4.0 M in dioxane, 1.10mL, 4.40 mL). The resulting mixture was heated to 50° C. for 2 h. Atcompletion, the reaction was concentrated, then azeotroped with tolueneto provide (S)-2-oxa-8-azaspiro[4.5]decan-4-amine LCMS: m/z 171 (M+H)⁺.

Intermediate 5 (R)-8-azaspiro[4.5]decan-1-amine

Intermediate 5 was synthesized using the above procedure ormodifications to the procedure applied in the synthesis of Intermediate4 using the corresponding ketone and sulfinamide.

Intermediate 6 (3 S,4S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate

(a) To a −10° C. solution of diisopropylamine (23.4 mL, 166 mmol) in THF(220 mL) was added n-BuLi (2.5 M in hexane, 64.1 mL, 160 mmol) dropwise.After stirring for 30 min at this temperature, 1-tert-butyl 4-ethylpiperidine-1,4-dicarboxylate (27.5 g, 107 mmol) in THF (50 mL) was addeddropwise and the resulting mixture was stirred for 30 min at 0° C.(S)-2-((tert-butyldimethylsilyl)oxy)propanal (20.47 mL, 102 mmol) wasadded and the mixture was stirred for 1 h at 0° C., then 1 h at roomtemperature. The reaction was diluted with sat. aq NaHCO₃:H₂O (1:4, 125mL), EtOAc (50 mL) was added, and the phases were separated. The aqueousphase was further extracted with EtOAc (3×100 mL). The combined organicphases were dried over Na₂SO₄, filtered, and the solvent was removedunder reduced pressure. The resulting residue was used in next stepwithout further purification. LCMS: m/z 346.4 (M+H-Boc)⁺.

(b) To a solution of crude 1-tert-butyl 4-ethyl4-((2S)-2-((tert-butyldimethylsilyl)oxy)-1-hydroxypropyl)piperidine-1,4-dicarboxylate(95 g, 214 mmol) in THF (600 mL) was added LiBH₄ (7.0 g, 321 mmol)portionwise. The resulting mixture was stirred for 16 h at roomtemperature. After cooling to 0° C., sat. aq NaHCO₃:H₂O (1:2, 150 mL)was added and the resulting mixture was vigorously stirred untilbubbling subsided. The mixture was diluted with EtOAc (100 mL) and themixture was filtered. The liquid phases were separated, and the aqueousphase was further extracted with EtOAc (3×50 mL). The combined organicphases were washed with brine, dried over Na₂SO₄, filtered, andconcentrated to give tert-butyl4-((2S)-2-((tert-butyldimethylsilyl)oxy)-1-hydroxypropyl)-4-(2-hydroxyethyl)piperidine-1-carboxylate(64.8 g, 161 mmol), which was used in next step without furtherpurification.

(c) A solution of tert-butyl4-((2S)-2-((tert-butyldimethylsilyl)oxy)-1-hydroxypropyl)-4-(2-hydroxyethyl)piperidine-1-carboxylate(64.8 g, 161 mmol) in THF (500 mL) was treated with TBAF (1 M in THF,242 mL, 242 mmol), then stirred for 2 h at room temperature. Atcompletion, the reaction mixture was treated with aq NaHCO₃:H₂O (1:2,150 mL) and the phases were separated. The aqueous phase was extractedwith EtOAc (3×100 mL). The combined organic phases were washed withbrine, dried over Na₂SO₄, filtered, and concentrated. The resultingresidue was purified by flash chromatography over silica gel (20-100%EtOAc/heptane eluent) to give tert-butyl4-((2S)-1,2-dihydroxypropyl)-4-(2-hydroxyethyl)piperidine-1-carboxylate(39.25 g, 136 mmol).

(d) To a 0° C. suspension of NaH (10.60 g, 424 mmol, 60 wt % in mineraloil) in THF (600 mL) was added dropwise a solution of tert-butyl4-((2S)-1,2-dihydroxypropyl)-4-(2-hydroxyethyl)piperidine-1-carboxylate(35.06 g, 121 mmol), followed by a solution of p-toluenesulfonylchloride (23.10 g, 121 mmol) in THF (200 mL). The resulting mixture wasstirred for 1 h at 0° C. At completion, sat. aq NH₄Cl (˜5 mL) was addedslowly at −20° C. and the reaction was vigorously stirred until bubblingceased. Then, sat. aq NH₄Cl (100 mL) and brine (100 mL) weresequentially added. The mixture was extracted with EtOAc (3×100 mL) andthe combined organic phases were dried over Na₂SO₄, filtered, andconcentrated give (3S)-tert-butyl4-hydroxy-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (32.19 g,119 mmol) which was used in next step without further purification.LCMS: m/z 171.1 (M-Boc)⁻.

(e) A solution of (3S)-tert-butyl4-hydroxy-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (32.19 g,119 mmol) and Dess-Martin periodinane (67.4 g, 154 mmol) in DCM (300 mL)was stirred for 2 h at 0° C. After warming to room temperature, thereaction was concentrated and the resulting residue was purified byflash chromatography over silica gel (0-40% EtOAc/heptane eluent) togive (S)-tert-butyl3-methyl-4-oxo-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (27.68 g, 92mmol) as a pale yellow oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.09(d, J=9.60 Hz, 1H), 3.66-3.86 (m, 4H), 3.03 (ddd, J=13.77, 9.73, 3.79Hz, 1H), 2.90 (ddd, J=13.64, 10.23, 3.41 Hz, 1H), 1.68 (ddd, J=13.83,9.92, 4.29 Hz, 1H), 1.41-1.59 (m, 2H), 1.30-1.40 (m, 10H), 1.20-1.25 (m,3H).

(f) A solution of (3S)-tert-butyl3-methyl-4-oxo-2-oxa-8-azaspiro[4.5]decane-8-carboxylate (22.52 g mg, 84mmol), titanium(IV) ethoxide (70.1 mL, 334 mmol), and(R)-2-methylpropane-2-sulfinamide (21 g, 173 mmol) in THF (300 mL) wasstirred for 21 h at 90° C. After cooling to −4° C., MeOH (30 mL) wasadded. Then, lithium borohydride (1.82 g, 84 mmol) was added carefullysuch that the reaction temperature was maintained below −2° C. Theresulting mixture was stirred for 1 h at −4° C. At completion, sat. aqNH₄Cl was slowly added to quench the excess borohydride and the reactionmixture was diluted with EtOAc (500 mL). The resulting mixture wasvigorously stirred for 15 min at room temperature and then filteredthrough a pad of Celite, rinsing with EtOAc (500 mL). The mixture wasconcentrated and the resulting residue was purified by flashchromatography over silica gel (0-100% EtOAc/heptane eluent) to give(3S,4S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylateas a 95:5 diastereomeric mixture (minor diastereomer (3R,4S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate).

(g) The diastereomers were separated by chiral SFC as follows: column:LC-4 30×250 mm, flow rate: 100 g per minute, mobil phase: 30% MeOH inCO₂, detection: 225 nm, R_(t): 0.95 min (minor diastereomer R_(t): 0.55min) to give (3S,4S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate(19 g, 50.68 mmol). LCMS: m/z 375.2.

(h) To a solution of (3S,4S)-tert-butyl4-((R)-1,1-dimethylethylsulfinamido)-3-methyl-2-oxa-8-azaspiro[4.5]decane-8-carboxylate(165 mg, 0.44 mmol) in MeOH (0.88 mL) was added HCl (4.0 M in dioxane,1.10 mL, 4.40 mmol). The reaction was maintained at 50° C. for 2 h. Atcompletion, the reaction was concentrated and dried under vacuum toprovide (3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine. LCMS: m/z171.2 (M+H)⁺.

Intermediate 8 (1R,3R)-tert-butyl3-((tert-butyldimethylsilyl)oxy)-1-((R)-1,1-dimethylethylsulfinamido)-8-azaspiro[4.5]decane-8-carboxylate)

(a) A mixture of CuCl (142 mg, 1.432 mmol), (S)-TolBINAP (972 mg, 1.432mmol), and sodium tert-butoxide (138 mg, 1.432 mmol) in THF (60 mL) wasstirred for 30 min at room temperature.4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (13.34 g,52.5 mmol) in THF (20 mL) was added and the resulting mixture wasstirred for 10 min at room temperature. A solution of tert-butyl1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate (12.0 g, 47.7 mmol) in THF(50 mL) was added followed by MeOH (3.9 mL, 95 mmol). The resultingmixture was stirred for 16 h at room temperature. H₂O (150 mL) was addedfollowed by sodium perborate (36.7 g, 239 mmol) and the resultingmixture was vigorously stirred for 1 h at room temperature. Theresulting green suspension was filtered through a pad of Celite, pouredinto a separation funnel containing sat. aq NaHCO₃: sat. aq Na₂SO₃ (1:1,300 mL) and extracted with EtOAc (4×40 mL). The combined organic phaseswere dried over MgSO₄, filtered, and the volatiles were removed underreduced pressure to give (R)-tert-butyl3-hydroxy-1-oxo-8-azaspiro[4.5]decane-8-carboxylate crude. Enantiomericdetermination of this mixture show 90% ee (R_(t)(S): 1.59 min, R_(t)(R):1.80 min; chiral SFC; column: IA 4.6×100 mm, flow rate: 70 g per minute,mobile phase: 5-55% MeOH in CO₂, detection: 220 nm UV).

(b) A mixture of crude (R)-tert-butyl3-hydroxy-1-oxo-8-azaspiro[4.5]decane-8-carboxylate (theor. 47.7 mmol),imidazole (4.87 g, 71.6 mmol), and tert-butylchlorodimethylsilane (8.99g, 59.6 mmol) in DMF (120 mL) was stirred for 16 h at room temperature.The reaction mixture was poured into a separation funnel containing sat.aq NH₄Cl:H₂O (1:1, 250 mL) and extracted with Et₂O (5×50 mL). Thecombined organic phases were dried over MgSO₄, filtered and thevolatiles were removed under reduced pressure. The resulting residue waspurified by silica chromatography (0-30% EtOAc/heptane eluent) to give(R)-tert-butyl3-((tert-butyldimethylsilyl)oxy)-1-oxo-8-azaspiro[4.5]decane-8-carboxylate(13.12 g, 34.2 mmol) as colorless oil that solidified upon standing.

(c) A solution of (R)-tert-butyl3-((tert-butyldimethylsilyl)oxy)-1-oxo-8-azaspiro[4.5]decane-8-carboxylate(8 g, 20.86 mmol), titanium(IV) ethoxide (17.49 mL, 83.0 mmol), and(R)-2-methylpropane-2-sulfinamide (5.06 g, 41.7 mmol) in THF (100 mL)was stirred at room temperature for 16 h at 65° C. After cooling to −78°C., MeOH (15 mL) was added followed by lithium borohydride (1.363 g,62.6 mmol). The resulting mixture was stirred for 16 h at −78° C. Atcompletion, sat. aq NH₄Cl was slowly added to quench the excess ofborohydride followed by addition of EtOAc (100 mL). The resultingmixture was vigorously stirred for 15 min and then filtered through apad of Celite. The volatiles were removed under reduced pressure and theresulting residue was purified by flash chromatography over silica gel(0-50% EtOAc/heptane eluent) to give (1R,3R)-tert-butyl3-((tert-butyldimethylsilyl)oxy)-1-((R)-1,1-dimethylethylsulfinamido)-8-azaspiro[4.5]decane-8-carboxylate(5.3 g, 10.84 mmol) as a white solid. LCMS: m/z 489.3 (M+H)⁺.

Intermediate 9 tert-butyl1-((tert-butoxycarbonyl)amino)-3,3-difluoro-8-azaspiro[4.5]decane-8-carboxylate

(a) A mixture of tert-butyl3-((tert-butyldimethylsilyl)oxy)-1-(1,1-dimethylethylsulfinamido)-8-azaspiro[4.5]decane-8-carboxylate(365 mg, 0.746 mmol) and HCl (4 M in dioxane, 1.86 mL, 7.46 mmol) inMeOH (4 mL) was stirred for 1 h at 40° C. After cooling to roomtemperature, the reaction was concentrate to provide a white solid.LCMS: m/z 171.1 (M+H)⁺. A mixture of this residue, DIPEA (2.6 mL, 14.92mmol), and Boc₂O (407 mg, 1.865 mmol) in THF (15 mL) was stirred for 16h at room temperature. The mixture was poured into a separation funnelcontaining sat. aq NH₄Cl and extracted with Et₂O (5×10 mL). The combinedorganic phases were dried over MgSO₄, filtered and concentrated. Theresulting residue was purified by silica chromatography (10-80%EtOAc/heptane eluent) to give tert-butyl1-((tert-butoxycarbonyl)amino)-3-hydroxy-8-azaspiro[4.5]decane-8-carboxylate(275 mg, 0.742 mmol). LCMS: m/z 271.3 (M+H-Boc)⁺.

(b) A mixture of tert-butyl1-((tert-butoxycarbonyl)amino)-3-hydroxy-8-azaspiro[4.5]decane-8-carboxylate(275 mg, 0.742 mmol) and Dess-Martin periodinane (472 mg, 1.113 mmol) inDCM (7.5 mL) was stirred for 2 h at 0° C. The mixture was poured into aseparation funnel containing sat. aq NaHCO₃ and extracted with DCM (3×10mL). The combined organic phases were dried over MgSO₄, filtered andconcentrated. The resulting residue was purified by silicachromatography (5-75% EtOAc/heptane eluent) to give tert-butyl1-((tert-butoxycarbonyl)amino)-3-oxo-8-azaspiro[4.5]decane-8-carboxylate(135 mg, 0.366 mmol). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.57 (d,J=9.09 Hz, 1H), 4.16 (d, J=8.08 Hz, 1H), 3.89-4.08 (m, 2H), 2.77-2.93(m, 2H), 2.71 (dd, J=18.95, 8.08 Hz, 1H), 2.50 (d, J=18.19 Hz, 1H),2.07-2.24 (m, 2H), 1.76 (td, J=12.82, 4.67 Hz, 1H), 1.58-1.70 (m, 1H),1.42-1.53 (m, 18H), 1.25-1.38 (m, 1H).

(c) A mixture of tert-butyl1-((tert-butoxycarbonyl)amino)-3-oxo-8-azaspiro[4.5]decane-8-carboxylate(95 mg, 0.258 mmol) and DeoxoFluor (190 μL, 1.031 mmol) in DCM (1 mL)was stirred for 48 h at 50° C. The mixture was poured into a separationfunnel containing sat. aq NaHCO₃/ice and extracted with EtOAc (3×5 mL).The combined organic phases were dried over MgSO₄, filtered andconcentrated. The resulting residue was purified by silicachromatography (0-30% EtOAc/heptane eluent) to give tert-butyl1-((tert-butoxycarbonyl)amino)-3,3-difluoro-8-azaspiro[4.5]decane-8-carboxylate(52 mg, 0.133 mmol). ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.55 (d,J=9.35 Hz, 1H), 3.78-4.02 (m, 3H), 2.64-2.86 (m, 2H), 2.38-2.59 (m, 1H),2.10-2.32 (m, 1H), 1.79-2.10 (m, 2H), 1.58 (qd, J=12.72, 3.79 Hz, 1H),1.27-1.52 (m, 21H).

(d) A solution of tert-butyl1-((tert-butoxycarbonyl)amino)-3,3-difluoro-8-azaspiro[4.5]decane-8-carboxylate(50 mg, 0.128 mmol) in DCM was treated with TFA (0.5 mL). The resultingmixture was stirred at room temperature for 2 h. At completion, thereaction was concentrated to provide3,3-difluoro-8-azaspiro[4.5]decan-1-amine, which was used directly.LCMS: m/z 191.2 (M+H)⁺.

Intermediate 10 (R)-3,3-difluoro-8-azaspiro[4.5]decan-1-amine

Intermediate 10 was synthesized by the procedure applied in thesynthesis of Intermediate 9 using the corresponding starting materials.

Intermediate 11 (1R,3S)-3-fluoro-8-azaspiro[4.5]decan-1-amine

(a) To a solution of tert-butyl(1R,3R)-1-(((R)-tert-butylsulfinyl)amino)-3-hydroxy-8-azaspiro[4.5]decane-8-carboxylate(400 mg, 1.07 mmol) in DCM (8.5 mL) was added diethylaminosulfurtrifluoride (1.97 mL, 1.75 mmol) dropwise. The resulting mixture wasstirred at 0° C. for 90 min. At completion, the reaction was treatedwith sat. aq NaHCO₃ and the mixture was stirred at 0° C. for 10 min. Themixture was transferred to a separatory funnel, the organics wereseparated and the aqueous phase was washed with DCM (3×10 mL). Thecombined organics were dried over MgSO₄, filtered and concentrated toprovide tert-butyl(1R,3S)-1-(((R)-tert-butylsulfinyl)amino)-3-fluoro-8-azaspiro[4.5]decane-8-carboxylate(402 mg, 1.07 mmol).

(b) To a solution of tert-butyl(1R,3S)-1-(((R)-tert-butylsulfinyl)amino)-3-fluoro-8-azaspiro[4.5]decane-8-carboxylate(402 mg, 1.07 mmol) in MeOH (4 mL) was added HCl (4.0 M in dioxane, 2.67mL, 10.68 mmol). The resulting mixture was warmed to 45° C. for 45 min.At completion, the reaction was concentrated to provide(1R,3S)-3-fluoro-8-azaspiro[4.5]decan-1-amine LCMS: m/z 173.2 (M+H)⁺.

Intermediate 12 3-azaspiro[5.5]undecan-7-amine

(a) A mixture of commercially available tert-butyl7-oxo-3-azaspiro[5.5]undecane-3-carboxylate (0.74 g, 2.77 mmol) and2-methylpropane-2-sulfinamide (0.67 g, 5.54 mmol) in THF (500 mL) wastreated with tetraethoxytitanium (2.32 mL, 11.07 mmol), then heated to75° C. for 20 h. The reaction was cooled to 0° C., treated with MeOH (3mL) and lithium tetrahydroborate (0.181 g, 8.30 mmol), then the coldbath was removed to allow the reaction to warm to room temperature.After 2 h, sat. aq NH₄Cl (1 mL) was added to the reaction mixture togenerate a gel, which was diluted with EtOAc (30 mL). The resultingsuspension was stirred at room temperature for 15 min, and then filteredthrough a plug of Celite, rinsing with EtOAc. The organics wereconcentrated, and the crude residue was purified by flash chromatographyover silica gel (5-95 EtOAc/heptane eluent) to provide tert-butyl7-(1,1-dimethylethylsulfinamido)-3-azaspiro[5.5]undecane-3-carboxylate(625 mg, 61% yield). LCMS: m/z 373 (M+H)⁺.

(b) A room temperature solution of tert-butyldimethylethylsulfinamido)-3-azaspiro[5.5]undecane-3-carboxylate (625 mg,1.68 mmol) in DCM (6 mL) was treated with HCl (4.0 N in dioxane, 4 mL).After 2 h, the reaction was concentrated to provide2-methyl-N-(3-azaspiro[5.5]undecan-7-yl)propane-2-sulfinamide, which wasused directly in the next reaction. LCMS: m/z 273 (M+H)⁺.

(c) A solution of2-methyl-N-(3-azaspiro[5.5]undecan-7-yl)propane-2-sulfinamide in MeOH (4mL) was treated with HCl (4.0 N in dioxane, 3.67 mL) and heated to 45°C. for 45 min. At completion, the reaction was concentrated, azeotropedwith DCE then dried under vacuum. LCMS: m/z 169 (M+H)⁺.

Intermediate 13 tert-Butyl((1R,3R)-3-(trifluoromethyl)-8-azaspiro[4.5]decan-1-yl)carbamate

(a) A solution of benzyl 1-oxo-8-azaspiro[4.5]dec-2-ene-8-carboxylate(3.05 g, 10.7 mmol) in THF (40 mL) was cooled to 0° C.Trimethyl(trifluoromethyl)silane (6.41 mL of a 2 M stock solution inTHF, 12.83 mmol), then tetrabutylammonium fluoride (0.214 mL of a 1 Mstock solution in THF, 0.214 mmol) were added, and the resultingsolution was stirred at 0° C. for 1.5 h. The reaction was quenched with2 M aqueous HCl (10 mL) and stirred at 0° C. for 1 h. The solution wasdiluted with sat. aq NH₄Cl (50 mL) and extracted with EtOAc (3×50 mL).The combined organic layers were dried over Na₂SO₄, filtered, andconcentrated. The resulting residue was purified by silicachromatography (0-50% EtOAc/heptane eluent) to give benzyl1-oxo-3-(trifluoromethyl)-8-azaspiro[4.5]decane-8-carboxylate as acolorless oil (2.22 g, 6.25 mmol, 58% yield). ¹H NMR (400 MHz,CHLOROFORM-d) δ ppm 7.45-7.31 (m, 5H), 5.16 (s, 2H), 3.84 (dd, J=8.9,5.1 Hz, 1H), 3.30 (ddd, J=13.5, 9.5, 3.4 Hz, 1H), 3.21 (ddd, J=13.5,9.8, 3.6 Hz, 1H), 3.03-2.87 (m, 1H), 2.66 (ddd, J=18.8, 8.4, 1.5 Hz,1H), 2.46 (dd, J=18.9, 10.7 Hz, 1H), 2.38-2.25 (m, 1H), 1.97-1.79 (m,2H), 1.70-1.58 (m, 1H), 1.54 (m, 3H). ¹⁹F NMR (376 MHz, CHLOROFORM-d) 6ppm-72.08 (d, J=8.0 Hz). ¹³C NMR (101 MHz, CHLOROFORM-d) δ ppm 215.93,155.18, 136.67, 128.53, 128.07, 127.93, 125.74 (q, J=263 Hz), 67.24,47.96, 40.35, 39.86, 37.30, 32.77 (q, J=29 Hz), 33.77 (q, J=3 Hz),31.89, 31.10.

(b) A solution of benzyl1-oxo-3-(trifluoromethyl)-8-azaspiro[4.5]decane-8-carboxylate (2.22 g,6.25 mmol), (R)-tert-butanesulfinamide (1.514 g, 12.50 mmol), andtetraethoxytitanium (5.70 g, 5.24 mL, 25.00 mmol) in 50 mL dry THF washeated to 80° C. for 16 h. The reaction was cooled to −78° C., then MeOH(10 mL) and lithium borohydride (0.408 g, 18.74 mmol) were added. Thereaction was allowed to warm to room temperature over 3 h. The reactionwas quenched by addition of sat. aq NH₄Cl (50 mL). The resultingheterogeneous mixture was filtered through Celite, rinsing with EtOAc.The layers of the filtrate were separated and the aqueous layer wasextracted with EtOAc (2×25 mL). The combined organic layers were driedover Na₂SO₄, filtered, and the volatiles were removed under reducedpressure. The resulting white solid, benzyl(1R)-1-(((R)-tert-butylsulfinyl)amino)-3-(trifluoromethyl)-8-azaspiro[4.5]decane-8-carboxylate,was used without further purification. LCMS: m/z 461.3 (M+H)⁺.

(c) To a solution of benzyl(1R)-1-(((R)-tert-butylsulfinyl)amino)-3-(trifluoromethyl)-8-azaspiro[4.5]decane-8-carboxylate(assumed 2.88 g, 6.25 mmol) in MeOH (25 mL) was added 4 M HCl in dioxane(3.13 mL, 12.51 mmol). The resulting solution was stirred at roomtemperature for 1 h. Volatiles were removed under reduced pressure andthe resulting residue was dried under vacuum for 2 h. The residue wasdissolved in DCM and N-ethyldiisopropylamine (5.57 mL, 31.3 mmol) anddi-tert-butyldicarbonate (2.05 g, 9.4 mmol) were added. The resultingmixture was stirred at ambient temperature for 72 h. The reaction wasquenched with sat. aq NH₄Cl (50 mL) and extracted with DCM (3×25 mL).The combined organic layers were dried over Na₂SO₄, filtered, and thevolatiles were removed under reduced pressure. The resulting residue waspurified by flash chromatography over silica gel (0-100% EtOAc/heptaneeluent) to afford diastereomerically pure benzyl(1R,3R)-1-((tert-butoxycarbonyl)amino)-3-(trifluoromethyl)-8-azaspiro[4.5]decane-8-carboxylate(0.90 g, 1.97 mmol, 31% yield). 41 NMR (400 MHz, CHLOROFORM-d) δ ppm7.34-7.20 (m, 5H), 5.06 (s, 2H), 4.39 (d, J=9.6 Hz, 1H), 3.96-3.75 (m,3H), 2.99 (t, J=11.0 Hz, 2H), 2.65 (dq, J=18.3, 9.1 Hz, 1H), 2.24 (dt,J=15.3, 8.3 Hz, 1H), 1.77 (dd, J=13.9, 9.7 Hz, 1H), 1.66 (dd, J=13.9,8.4 Hz, 1H), 1.60-1.41 (m, 3H), 1.39 (s, 9H), 1.31-1.16 (m, 2H). Amixture of diastereomers (1.47 g, 51% yield) was also recovered.

(d) A solution of benzyl(1R,3R)-1-((tert-butoxycarbonyl)amino)-3-(trifluoromethyl)-8-azaspiro[4.5]decane-8-carboxylate(0.90 g, 1.97 mmol) and 10% Pd/C (200 mg) in EtOH (40 mL) was stirredfor 2 h under 1 atm of hydrogen gas. At this time, the mixture wassparged with nitrogen for 5 min, then filtered through Celite under ablanket of nitrogen, rinsing the filter cake with EtOH. The filtrate wasconcentrated under reduced pressure and dried under vacuum to givetert-butyl((1R,3R)-3-(trifluoromethyl)-8-azaspiro[4.5]decan-1-yl)carbamate (625mg, 1.94 mmol, 98% yield) as a white foam. 41 NMR (400 MHz,CHLOROFORM-d) δ ppm 4.54 (d, J=9.7 Hz, 1H), 3.84 (q, J=8.8 Hz, 1H), 3.00(tt, J=12.1, 4.0 Hz, 2H), 2.79-2.63 (m, 3H), 2.28 (ddd, J=13.5, 8.8, 6.8Hz, 1H), 2.19 (d, J=8.5 Hz, 1H), 1.80 (qd, J=14.0, 9.1 Hz, 2H), 1.63(qd, J=9.0, 3.4 Hz, 2H), 1.47 (m, 12H).

Example 16-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloropyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one

General Synthetic Route Method A

(a) Hydrazine hydrate (9.17 mL, 187 mmol) was added to a solution of6-chloro-3-methylpyrimidine-2,4(1H,3H)-dione (10 g, 62.3 mmol) in EtOH(200 mL). The resulting mixture was stirred at 80° C. The slurry becameincreasingly thick over the course of 5-10 min. The heating wascontinued for 1 h, then the heat bath was removed and the reactionmixture was allowed to cool to room temperature for 1 h. The resultingsuspension was filtered and the filter cake was washed with EtOH (100mL), then dried under vacuum. The intermediate6-hydrazinyl-3-methylpyrimidine-2,4(1H,3H)-dione was obtained as a whitepowder (9.7 g, 100% yield) and was used directly in the nexttransformation.

(b) To a suspension of 6-hydrazinyl-3-methylpyrimidine-2,4(1H,3H)-dione(5.26 g, 33.7 mmol) in MeOH (25 mL) was added 4-methoxybenzaldehyde(5.50 g, 40.4 mmol). The resulting mixture was stirred at roomtemperature for 30 min. The slurry became very thick, and was dilutedwith MeOH (25 mL) to facilitate stirring. The reaction was maintainedfor an additional 30 min. The slurry was filtered and the filter cakewas dried under vacuum for 16 h to provide6-((4-methoxybenzyl)diazenyl)-3-methylpyrimidine-2,4(1H,3H)-dione as awhite solid (8.0 g, 87% yield). LCMS: m/z 275 (M+H)⁺, R_(t)=0.43 min.

(c) To a suspension of6-((4-methoxybenzyl)diazenyl)-3-methylpyrimidine-2,4(1H,3H)-dione (0.68g, 2.479 mmol) in 2:1 DMF:i-PrOH (12 mL) was added2,3-dichloroisonicotinaldehyde (0.436 g, 2.479 mmol) and piperidine(0.241 mL, 2.430 mmol). The resulting mixture was stirred at 85° C. for1 h. At completion, the reaction mixture was partitioned between water(50 mL) and EtOAc (50 mL). The layers were separated and the aqueouslayer was washed with EtOAc (50 mL). The combined organics were washedwith brine (40 mL), dried with Na₂SO₄, filtered and concentrated. Thecrude was purified by flash chromatography over silica gel (0-10%MeOH/DCM eluent) to afford3(2,3-dichloropyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidine-4,6(5H,7H)-dioneas a yellow solid (420 mg, 39% yield, 90% purity). LCMS: m/z 432 (M+H)⁺,R_(t)=1.29 min.

(d) To a suspension of3-(2,3-dichloropyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidine-4,6(5H,7H)-dione(133 mg, 0.308 mmol) in DMF (1.5 mL), was added(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(408 mg, 0.923 mmol). The reaction was maintained for 5 min before theaddition of (3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine (112 mg,0.462 mmol) and DBU (278 μL, 1.846 mmol). The reaction was stirred atroom temperature for 1.5 h. At completion, the reaction mixture waspartitioned between water (20 mL) and EtOAc (20 mL). The layers wereseparated and the aqueous layer was washed with EtOAc (20 mL). Thecombined organics were washed with brine (15 mL), dried over Na₂SO₄,filtered and concentrated. The crude was purified by flashchromatography over silica gel (0-10% MeOH/DCM eluent) to provide6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloropyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one(140 mg, 70% yield, 90% purity). LCMS: m/z 584 (M+H)⁺, R_(t)=1.04 min.

(e) A solution of6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloropyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one(56 mg, 0.099 mmol) in DCM (493 uL, 0.2 M) was treated with TFA (449 mg,3.94 mmol) and triflic acid (8.75 μL, 0.099 mmol). The reaction wasmaintained for 10 min, at which point complete conversion was observedby LCMS. The reaction mixture was concentrated, and the resultingresidue was azeotroped with toluene (5 mL). The crude was purified byprep HPLC to provide6-43S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloropyridin-4-yl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one(12.0 mg, 26% yield). LCMS: m/z 464 (M+H)⁺, R_(t)=0.89 min; ¹H NMR (400MHz, METHANOL-d₄) δ ppm 8.37 (d, J=4.8 Hz, 1H), 7.53 (d, J=4.8 Hz, 1H),3.42-3.65 (m, 5H), 2.98-3.17 (m, 2H), 2.86 (t, J=7.3 Hz, 1H), 1.98-2.20(m, 1H), 1.25-1.93 (m, 9H). IC₅₀=0.028 μM.

Example 2(R)-6-(1-amino-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichlorophenyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one

General Synthetic Route Method B

tert-butyl(R)-(8-(2-benzyl-3-(2,3-dichlorophenyl)-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-8-azaspiro[4.5]decan-1-yl)carbamatewas prepared by General Method A, substituting benzyaldehyde forp-methoxybenzaldehyde in step b, 2,3-dichlorobenzaldehyde for2,3-dichloroisonicotinaldehyde in step c and (R)-tert-butyl8-azaspiro[4.5]decan-1-ylcarbamate for(3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine in step d.

The synthesis concludes as follows: (a) To a solution of tert-butyl41R)-8-(2-benzyl-3-(2,3-dichlorophenyl)-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-8-azaspiro[4.5]decan-1-yl)carbamate(0.889 g, 1.394 mmol) in DMSO (14 mL), sodium tert-butoxide (1.340 g,13.94 mmol) was added. A balloon of oxygen was bubbled through thereaction mixture with stirring for 15 min. At completion, the reactionmixture was partitioned between water (50 mL) and EtOAc (50 mL). Thelayers were separated and the aqueous layer was extracted with EtOAc (50mL). The organics were combined, dried with Na₂SO₄, filtered andconcentrated. The crude was purified by flash chromatography over silicagel (0-10% MeOH/DCM eluent) to provide (R)-tert-butyl(8-(3-(2,3-dichlorophenyl)-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-8-azaspiro[4.5]decan-1-yl).LCMS: m/z 547 (M+H)⁺, R_(t)=1.57 min.

(b) To a solution of (R)-tert-butyl(8-(3-(2,3-dichlorophenyl)-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-8-azaspiro[4.5]decan-1-yl)carbamatein DCM was added HCl (4.0 M in dioxane, 1.827 mL, 7.31 mmol). Theresulting mixture was stirred at room temperature for 1 h, thenconcentrated. The crude was purified by prep HPLC to afford(R)-6-(1-amino-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichlorophenyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one(50 mg, 0.111 mmol, 15% yield, 2 steps) as a white solid followinglyophilization of pure fractions. LCMS: m/z 447 (M+H)⁺, R_(t)=0.97 min;¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.63 (dd, J=7.8, 1.8 Hz, 1H),7.29-7.47 (m, 2H), 3.43-3.57 (m, 5H), 2.98-3.14 (m, 2H), 2.87 (t, J=7.3Hz, 1H), 2.05 (dd, J=7.2, 4.2 Hz, 1H), 1.35-1.95 (m, 9H). IC₅₀=0.0255μM.

Example 32-(4-(aminomethyl)-4-methylpiperidin-1-yl)-5-(2,3-dichlorophenyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one

General Synthetic Route Method C

(a) A solution of 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine (2.5 g, 13.30mmol) and 4-methoxybenzyl alcohol (1.82 mL, 14.63 mmol) in 1,4-dioxane(33.2 mL) was treated with potassium tert-butoxide (5.97 g, 53.2 mmol),and the reaction mixture was stirred at room temperature for 3 h. Atcompletion, the reaction mixture was transferred to a separatory funnelcontaining sat. aq NH₄Cl (150 mL, 1:1 sat. NH₄Cl to deionized water),and extracted with DCM (4×150 mL). The combined extracts were dried overMgSO₄, filtered, and concentrated to afford2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine as anorange solid. LCMS: m/z 290 (M+H)⁺, R_(t)=1.44 min; ¹H NMR (400 MHz,DMSO-d₆) δ ppm 12.23 (br. s., 1H), 7.46 (d, J=8.34 Hz, 2H), 7.35-7.39(m, 1H), 6.93-6.99 (m, 2H), 6.49 (dt, J=2.27, 1.14 Hz, 1H), 5.46 (s,2H), 3.72-3.78 (m, 3H).

(b) To a −20° C. solution of2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine (1.50 g,5.18 mmol) in DMF (13 mL) was added N-bromosuccinimide (1.014 g, 5.70mmol). After stirring for 10 min, LCMS indicated complete consumption ofstarting material. The reaction mixture was transferred to a separatoryfunnel containing sat. aq Na₂SO₃ (150 mL) and extracted with Et₂O (4×150mL). The combined extracts were dried over MgSO₄, filtered, andconcentrated to afford5-bromo-2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine asan off-white solid (1.89 g, 90% yield). LCMS: m/z 368 (M+H)⁺, R_(t)=1.56min; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.60 (d, J=2.53 Hz, 1H), 7.48(d, J=8.59 Hz, 2H), 6.97 (d, J=8.84 Hz, 2H), 5.49 (s, 2H), 3.76 (s, 3H).

(c) To a stirred solution of5-bromo-2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine(600 mgs, 1.481 mmol), DIPEA (0.517 mL, 2.96 mmol), and DMAP (9.05 mg,0.074 mmol) in THF (8.7 mL) was added di-tert-butyl dicarbonate (0.516mL, 2.222 mmol), and the reaction was stirred for 30 min. At completion,the reaction mixture was diluted with EtOAc (5 mL), transferred to aseparatory funnel containing sat. aq NH₄Cl (15 mL) and extracted withEtOAc (3×15 mL). The combined extracts were dried over MgSO₄, filteredand concentrated to afford tert-butyl5-bromo-2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylateas a pale orange solid (873 mg). This crude product was used in thesubsequent reaction without further purification. LCMS: m/z 490 (M+Na)⁺,R_(t)=1.58 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.89 (s, 1H), 7.49 (m,J=8.84 Hz, 2H), 6.98 (m, J=8.84 Hz, 2H), 5.51 (s, 2H), 3.77 (s, 3H),1.60 (s, 9H).

(d) A mixture of tert-butyl5-bromo-2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate(253 mgs, 0.54 mmol), (2,3-dichlorophenyl)boronic acid (129 mg, 0.675mmol), potassium phosphate (344 mg, 1.619 mmol), and PdCl₂(dppf)-DCMadduct (22 mg, 0.27 mmol) in THF:H₂O (10:1, 0.3 mL) was degassed, andthe mixture was microwave heated to 90° C. for 90 min LCMS analysisindicated complete consumption of starting material. The reactionmixture was transferred to a separatory funnel containing sat. aq NH₄Cl(10 mL) and extracted with EtOAc (3×10 mL). The combined extracts weredried over MgSO₄, filtered and concentrated to afford a black oil. Thecrude product was purified by flash chromatography over silica gel(0-40% EtOAc/heptane eluent). Concentration of pure fractions provided2-chloro-5-(2,3-dichlorophenyl)-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidineas a white solid (107 mgs, 37% yield). LCMS: m/z 556 (M+Na)⁺, R_(t)=1.71min; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.46-7.51 (m, 2H), 1.71 (s,9H), 7.26 (dd, J=7.58, 1.77 Hz, 1H), 7.18 (t, J=7.83 Hz, 1H), 7.13 (d,J=8.59 Hz, 2H), 6.78-6.89 (m, 2H), 5.38 (s, 2H), 3.83 (s, 3H).

(e) To a solution of2-chloro-5-(2,3-dichlorophenyl)-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine(125 mg, 0.201 mmol) in DIPEA (1 mL) and DMSO (1 mL) was addedtert-butyl ((4-methylpiperidin-4-yl)methyl)carbamate (50.5 mg, 0.221mmol), and the reaction mixture was heated to 90° C. for 4 h. Thereaction was treated with additional DIPEA (1.5 mL), DMSO (0.5 mL) andtert-butyl ((4-methylpiperidin-4-yl)methyl)carbamate (30 mg, 0.131 mmol)and heated at 90° C. for 16, then 130° C. for 2 h. The reaction wascooled and concentrated. The crude mixture was treated with DCM (2.5 mL)and TFA (1.0 mL) and maintained at room temperature with stirring for 3h. At completion, the reaction was concentrated. The liquor wasfree-based by filtration through StratoSpeheres SPE PL-HCO₃ MP-Resins,then purified by prep HPLC to provide2-(4-(aminomethyl)-4-methylpiperidin-1-yl)-5-(2,3-dichlorophenyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(2.7 mg, 3.3% yield). LCMS: m/z 406 (M+H)⁺, R_(t)=0.71 min; ¹H NMR (400MHz, METHANOL-d₄) δ ppm 7.41 (ddd, J=9.54, 7.89, 1.52 Hz, 2H), 7.19-7.31(m, 1H), 6.83 (s, 1H), 3.87 (dt, J=13.39, 4.42 Hz, 2H), 3.40 (ddd,J=13.52, 9.85, 3.41 Hz, 2H), 2.63 (s, 2H), 1.52-1.66 (m, 2H), 1.40-1.52(m, 2H), 1.08 (s, 3H). IC₅₀=0.062 μM.

Example 46-(4-amino-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one

2-benzyl-3-(2,3-dichlorophenyl)-6-hydroxy-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-onewas synthesized by General Method A, substituting benzyaldehyde forp-methoxybenzaldehyde in step b, and 2,3-dichlorobenzaldehyde for2,3-dichloroisonicotinaldehyde in step c.

The synthesis continued as follows: step a) A mixture of2-benzyl-3-(2,3-dichlorophenyl)-6-hydroxy-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one(1.77 g, 4.41 mmol) in phosphorous oxychloride (20.29 g, 132.0 mmol) washeated to 100° C. for 16 h. Then, tetramethylammonium chloride (1.45 g,13.23 mmol) was added and the reaction mixture was heated to 100° C. for24 h. The reaction was concentrated and treated with sat. aq NaHCO₃solution. The mixture was stirred vigorously, then the solids werefiltered and washed with water, and dried under vacuum. The crudeproduct was purified by flash chromatography (0-100% EtOAc/heptanedeluent) to provide2-benzyl-6-chloro-3-(2,3-dichlorophenyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-oneas a white solid (1.37 g, 92% yield). LCMS: m/z 419 (M+H)⁺, R_(t)=1.48min; ¹H NMR (400 MHz, DCM-d₂) δ 7.70 (dd, J=8.1, 1.6 Hz, 1H), 7.43-7.34(m, 1H), 7.33-7.26 (m, 3H), 7.21 (dd, J=7.7, 1.5 Hz, 1H), 7.12-7.01 (m,2H), 5.38 (d, J=10.7 Hz, 1H), 5.21 (d, J=14.9 Hz, 1H), 3.62 (s, 3H).

(b) A mixture of2-benzyl-6-chloro-3-(2,3-dichlorophenyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-one(100 mg, 0.143 mmol), tert-butyl (4-methylpiperidin-4-yl)carbamate (59.7mg, 0.279 mmol) and DIPEA (83 mg, 0.642 mmol) in NMP (1 mL) wasmicrowave heated at 120° C. for 90 min. At completion, the reaction wastreated with EtOAc and 10% citric acid. The layers were separated andthe aqueous layer was washed with EtOAc. The combined organics werewashed with 1:1 water:brine, dried over MgSO₄ and concentrated to give ayellow solid. The crude was purified by flash chromatography over silicagel (30-45% EtOAc/heptane eluent) to provide semi-pure tert-butyl(1-(2-benzyl-3-(2,3-dichlorophenyl)-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-4-methylpiperidin-4-yl)carbamate,which was used directly. LCMS: m/z 597 (M+H)⁺, R_(t)=1.70 min.

(c) The semi-pure tert-butyl(1-(2-benzyl-3-(2,3-dichlorophenyl)-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-4-methylpiperidin-4-yl)carbamatewas converted to6-(4-amino-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)-5-methyl-2H-pyrazolo[3,4-d]pyrimidin-4(5H)-oneby General Method B. LCMS: m/z 407 (M+H)⁺, R_(t)=0.88 min; ¹H NMR (400MHz, METHANOL-d₄) δ ppm 7.62 (dd, J=7.96, 1.64 Hz, 1H), 7.40-7.44 (m,1H), 7.33-7.39 (m, 1H), 3.48 (s, 3H), 3.34-3.43 (m, 2H), 3.22-3.29 (m,2H), 1.67-1.80 (m, 4H), 1.24 (s, 3H). IC₅₀=0.064 μM.

Example 52-(4-(aminomethyl)-4-methylcyclohexyl)-5-(2,3-dichlorophenyl)-3-methyl-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one

(a) To a room temperature solution of2-(methylthio)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one (2.0 g, 11.04 mmol)in 1 M aq NaOH (60 mL) was added dimethyl sulfate (6.96 g, 55.2 mmol).The resulting mixture was stirred at room temperature for 6 h. Atcompletion, the reaction was filtered and resulting the pale brown solidwas washed with water then dried under high vacuum for 3 h to afford3-methyl-2-(methylthio)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one (1.42 g,66% yield), which was used directly in the next reaction.

(b) To a 0° C. solution of3-methyl-2-(methylthio)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one (1.42 g,7.27 mmol) in DMF (20 mL) was added NaH (0.58 g, 14.55 mmol, 60% inmineral oil) portionwise over 5 min. The reaction was stirred at roomtemperature for 2 h, then cooled to 0° C. and treated with(2-(chloromethoxy)ethyl)trimethylsilane (1.27 g, 7.64 mmol). Theresulting mixture was stirred at 0° C. for 5 min, then the cold bath wasremoved and the mixture was stirred for 3 h. At completion, the reactionmixture was quenched by slow addition of sat. aq NH₄Cl, then dilutedwith Et₂O. The layers were separated and the organics were washed withsat. aq NH₄Cl, then dried over MgSO₄ and concentrated. The crudematerial was purified by flash chromatograpy over silica gel (0-50%EtOAc/heptane eluent) to provide3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(1.76 g, 74% yield). LCMS: m/z 326 (M+H)⁺, R_(t)=1.29 min; ¹H NMR (400MHz, CHLOROFORM-d) δ ppm 6.91 (d, J=3.54 Hz, 1H), 6.70 (d, J=3.54 Hz,1H), 5.52 (s, 2H), 3.64 (s, 3H), 3.52-3.61 (m, 2H), 2.67 (s, 3H),0.87-1.03 (m, 2H), 0.00 (d, J=6.57 Hz, 9H).

(c) To a −20° C. solution of3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(1.76 g, 5.41 mmol) in DMF (15 mL) was added NBS (1.01 g, 5.86 mmol).After 5 min, the reaction was complete. The reaction mixture wasquenched by addition of sat. aq Na₂S₂O₃, diluted with water, then twiceextracted with diethyl ether. The combined organics were dried overMgSO₄, filtered and concentrated. The resulting residue was purified byflash chromatography over silica gel (0-50% EtOAc/heptane eluent) toafford6-bromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-oneas a colorless oil (2.19 g, 100% yield). 41 NMR (400 MHz, CHLOROFORM-d)δ ppm 6.75 (s, 1H), 5.56-5.62 (m, 2H), 3.54-3.71 (m, 5H), 2.57-2.73 (m,3H), 0.87-1.03 (m, 2H), −0.04-0.03 (m, 9H); ¹³C NMR (101 MHz,CHLOROFORM-d) δ ppm 159.33 (s, 1 C), 159.05 (s, 1 C), 148.97 (s, 1 C),107.74 (s, 1 C), 107.10 (s, 1 C), 105.95 (s, 1 C), 78.80 (s, 1 C), 78.49(s, 1 C), 78.17 (s, 1 C), 73.70 (s, 1 C), 72.97 (s, 1 C), 68.27 (s, 1C), 68.05 (s, 1 C), 54.90 (s, 1 C), 31.44 (s, 1 C), 31.30 (s, 1 C),19.15 (s, 1 C), 16.73 (s, 1 C), 16.66 (s, 1 C), 0.25 (s, 1 C).

(d) To a room temperature solution of6-bromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(1.76 g, 4.35 mmol) in DMF (15 mL) was added NBS (968 mg, 5.43 mmol).The resulting mixture was stirred at room temperature for 30 min. Atcompletion, the reaction was quenched by addition of sat. aq Na₂S₂O₃,diluted with water, then twice extracted with Et₂O. The combinedorganics were dried over MgSO₄, filtered and concentrated to provide5,6-dibromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(1.82 g, 87% yield). The resulting material was used directly in thenext reaction. LCMS: m/z 484 (M+H)⁺, R_(t)=1.84 min.

(e) To a −78° C. solution of5,6-dibromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(1.82 g, 3.77 mmol) in THF (10 mL) was added dropwise a 1.7 M solutionof t-BuLi in THF (5.65 mmol, 3.32 mL). The reaction was maintained for 5min, then quenched with sat. aq NH₄Cl and the reaction was allowed towarm to room temperature. The reaction was partitioned between water andEtOAc. The organics were dried over MgSO₄ and concentrated, thenpurified by flash chromatograpy over silica gel (0-50% EtOAc/heptaneeluent) to provide5-bromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(820 mg, 54% yield) as a colorless oil that solidified upon standing.LCMS: m/z 404 (M+H)⁺, R_(t)=1.42 min; ¹H NMR (400 MHz, CHLOROFORM-d) δppm 6.92 (s, 1H), 5.46 (s, 2H), 3.50-3.64 (m, 5H), 2.64 (s, 3H),0.88-1.01 (m, 2H), −0.08-0.09 (m, 9H).

(f) To a room temperature solution of5-bromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(340 mg, 0.723 mmol) in EtOH (14.45 mL) was added Mo₇O₂₄(NH₄)₆—H₂O (268mg, 0.217 mmol) and 30% aq hydrogen peroxide (0.738 mL, 7.23 mmol) andthe resulting mixture was stirred at room temperature for 16 h. Atcompletion, the reaction was quenched by addition of sat. aq Na₂S₂O₃.The resulting suspension was stirred at room temperature for 5 min thenpoured into a separation funnel containing brine. The mixture wasextracted with EtOAc (5×5 mL). The combined organic phases were driedover MgSO₄, filtered and concentrated to afford semi-pure5-bromo-3-methyl-2-(methylsulfonyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(354 mg) which was used in the next step without purification. LCMS: m/z436 (M+H)⁺, R_(t)=1.62 min.

(g) A mixture of5-bromo-3-methyl-2-(methylsulfonyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one(679 mg, 0.723 mmol) and tert-butyl((4-methylpiperidin-4-yl)methyl)carbamate (165 mg, 0.723 mmol) in DIPEA(5 mL) was heated to 90° C. for 2 h. At completion, the reaction wasconcentrated and purified by flash chromatography over silica gel (0-40%EtOAc/heptane eluent) to provide tert-butyl((1-(5-bromo-3-methyl-4-oxo-7-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate(42 mg, 9% yield). LCMS: m/z 584 (M+H)⁺, R_(t)=1.49 min.

(h) To a mixture of tert-butyl((1-(5-bromo-3-methyl-4-oxo-7-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate(200 mg, 0.342 mmol) in 9:1 THF:H₂O (3 mL) was added(2,3-dichlorophenyl)boronic acid (82 mg, 0.428 mmol), potassiumphosphate (218 mg, 1.026 mmol), andchloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(13.5 mg, 0.017 mmol) under nitrogen. The mixture was microwave heatedto 100° C. for 60 min. At completion, the reaction was concentrated andthe resulting residue was purified by flash chromatograpy over silicagel (0-50% EtOAc/heptane) to provide semi-pure tert-butyl41-(5-(2,3-dichlorophenyl)-3-methyl-4-oxo-7-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate(138 mg, 55% yield). LCMS: m/z 650 (M+H)⁺, R_(t)=1.73 min.

(i) To a solution of tert-butyl41-(5-(2,3-dichlorophenyl)-3-methyl-4-oxo-7-((2-(trimethylsilyl)ethoxy)methyl)-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-2-yl)-4-methylpiperidin-4-yl)methyl)carbamate(138 mg, 0.212 mmol) in DCM (3 mL) was added TFA (1 mL). The reactionwas maintained at room temperature for 1 h. The reaction wasconcentrated and the resulting residue was dissolved in MeOH (2 mL) andtreated with ethylenediamine (191 mg, 3.18 mmol). The resulting mixturewas stirred at room temperature for 2 h. At completion, the reaction wasconcentrated and the resulting residue was purified by reverse phaseprep HPLC to provide2-(4-(aminomethyl)-4-methylpiperidin-1-yl)-5-(2,3-dichlorophenyl)-3-methyl-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-oneas a white powder (35 mg, 38% yield). LCMS: m/z 420 (M+H)⁺, R_(t)=0.79min; ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.41-7.49 (m, 1H), 7.37 (d,J=7.53 Hz, 1H), 7.24 (t, J=7.91 Hz, 1H), 6.95 (s, 1H), 3.50 (s, 3H),3.19-3.29 (m, 2H), 3.02-3.17 (m, 2H), 2.59 (s, 2H), 1.60-1.76 (m, 2H),1.52 (d, J=13.80 Hz, 2H), 1.05 (s, 3H). IC₅₀=0.100 μM.

Example 63-(6-amino-2,3-dichloropyridin-4-yl)-6-43S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one

(a) To a stirred, room temperature slurry of6-chloro-3-methylpyrimidine-2,4(1H,3H)-dione (3.6 g, 22.42 mmol) andpotassium carbonate (7.75 g, 56.1 mmol) in DMF (50 mL) was added1-(chloromethyl)-4-methoxybenzene (3.69 g, 23.54 mmol). The reaction waswarmed to 50° C. for 16 h, then cooled to room temperature andpartitioned between water and EtOAc. The organics were washed with waterand brine, then dried over MgSO₄ and concentrated. The crude waspurified by flash chromatography over silica gel (10-50% EtOAc/heptaneeluent) to provide6-chloro-1-(4-methoxybenzyl)-3-methylpyrimidine-2,4(1H,3H)-dione (4.4 g,70% yield). ¹H NMR (400 MHz, DMSO-d₆) δ ppm 7.21-7.28 (m, 2H), 6.91 (d,J=7.55 Hz, 2H), 6.12 (s, 1H), 5.14 (s, 2H), 3.68-3.79 (m, 3H), 3.13-3.20(m, 3H).

(b) To a solution of6-chloro-1-(4-methoxybenzyl)-3-methylpyrimidine-2,4(1H,3H)-dione (4.18g, 14.89 mmol) in EtOH (40 mL) was added hydrazine monohydrate (2.24 g,44.7 mmol). The mixture was heated at 80° C. for 2 h, then cooled toroom temperature and water (100 mL) was added. The solids were collectedby filtration, then dried under vacuum to afford6-hydrazinyl-1-(4-methoxybenzyl)-3-methylpyrimidine-2,4(1H,3H)-dione(3.52 g, 86% yield) as a white powder. LCMS: m/z 277 (M+H)⁺, R_(t)=0.47min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.05 (s, 1H), 7.09-7.24 (m, 2H),6.78-6.93 (m, 2H), 5.09-5.19 (m, 1H), 4.92-5.05 (m, 2H), 3.66-3.78 (m,4H), 3.07-3.19 (m, 3H).

(c) To a −40° C. solution6-hydrazinyl-1-(4-methoxybenzyl)-3-methylpyrimidine-2,4(1H,3H)-dione(3.52 g, 12.74 mmol) in DMF (100 mL) was added phosphoryl trichloride(14.25 mL, 153.0 mmol). The mixture was maintained at −40° C. for 2 h,then the cold bath was removed and the reaction was stirred at ambienttemperature for 10 h. At this time, the reaction was partiallyconcentrated under vacuum to remove excess phosphoryl trichloride, thenthe reaction mixture was cooled to 0° C. and water (200 mL) was added.The resulting precipitate was collected by vacuum filtration. The filtercake was rinsed with water, then dried under vacuum to provide7-(4-methoxybenzyl)-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(3.52 g, 100% yield) as a white solid. LCMS: m/z 287 (M+H)⁺, R_(t)=0.51min.

(d) A solution of7-(4-methoxybenzyl)-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(3.8 g, 13.27 mmol) in DMF (28 mL) was treated with N-iodosuccinimide(3.28 g, 14.60 mmol). At 2 h, incomplete conversion was observed. Thehot solution was treated with additional N-iodosuccinimide (1.1 g, 4.87mmol). At completion, the reaction was cooled to room temperature, thentreated with (1:1) sat. aq Na₂S₂O₃: sat. aq NaHCO₃ (50 mL) and stirredvigorously. The resulting mixture was diluted with water and theresulting precipitate was filtered and dried under vacuum to provide3-iodo-7-(4-methoxybenzyl)-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-c/]pyrimidine-4,6(5H)-dione(2.9 g, 53% yield). LCMS: m/z 413 (M+H)⁺, R_(t)=0.41 min.

(e) A slurry of3-iodo-7-(4-methoxybenzyl)-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-c/]pyrimidine-4,6(5H)-dione(740 mg, 1.80 mmol) and potassium carbonate (1.24 g, 8.98 mmol) inacetone (10 mL) was treated with benzyl bromide (338 mg, 1.98 mmol) andthe mixture was heated to 55° C. At completion, the reaction wasfiltered and concentrated, then purified by flash chromatography [0-5%MeOH/DCM eluent] to provide1-benzyl-3-iodo-7-(4-methoxybenzyl)-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-c/]pyrimidine-4,6(5H)-dione(840 mg, 1.67 g, 93% yield). LCMS: m/z 503 (M+H)⁺, R_(t)=0.96 min.

(f) A degassed mixture of1-benzyl-3-iodo-7-(4-methoxybenzyl)-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(770 mg, 1.53 mmol),(6-((tert-butoxycarbonyl)amino)-2,3-dichloropyridin-4-yl)boronic acid(1.18 g, 3.83 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]palladium(II)dichloride-DCM (125 mg, 0.153 mmol) and potassium carbonate (847 mg,6.13 mmol) in THF (10 mL) and water (2 mL) was heated to 70° C. After 30min, the reaction was complete by LCMS analysis. The reaction waspartitioned between water and EtOAc. The organics were washed withbrine, dried over MgSO₄ and concentrated. The crude was purified byflash chromatography (10-40% EtOAc/heptane eluent) to provide tert-butyl(4-(1-benzyl-7-(4-methoxybenzyl)-5-methyl-4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5,6-dichloropyridin-2-yl)carbamate(650 mg, 67% yield). LCMS: m/z 637 (M+H)⁺, R_(t)=0.93 min.

(g) A solution of tert-butyl(4-(1-benzyl-7-(4-methoxybenzyl)-5-methyl-4,6-dioxo-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-d]pyrimidin-3-yl)-5,6-dichloropyridin-2-yl)carbamate(440 mg, 0.69 mmol) in TFA (15 mL) was treated with triflic acid (311mg, 2.07 mmol) then heated to 70° C. After 1 h, the reaction wascomplete by LCMS analysis. The reaction was diluted with1,2-dichloroethane (20 mL), then concentrated to 10 mL volume. Theremaining acid was quenched with 5 N aq NaOH solution, adjusting topH-8. The mixture was then partitioned between water and EtOAc. Theorganics were washed with brine, dried over MgSO₄ and concentrated. Thecrude was purified by flash chromatography [0-10% MeOH/DCM eluent] toprovide3-(6-amino-2,3-dichloropyridin-4-yl)-1-benzyl-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(90 mg, 31% yield). LCMS: m/z 417 (M+H)⁺, R_(t)=0.69 min.

(h) To a vial charged with3-(6-amino-2,3-dichloropyridin-4-yl)-1-benzyl-5-methyl-1,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(90 mg, 0.216 mmol) was added(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphateand DMF (1 mL). The reaction was maintained with stirring for 15 minbefore the consecutive addition of(3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine-2HCl (73 mg, 0.302mmol) and DBU (230 mg, 1.51 mmol). After 16 h, the reaction was completeby LCMS analysis. The reaction was partitioned between water and EtOAc.The organics were washed with water and brine, dried over MgSO₄ andconcentrated. The crude was purified by flash chromatograpy (0-10%MeOH/DCM eluent) to provide3-(6-amino-2,3-dichloropyridin-4-yl)-6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-1-benzyl-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one(67 mg, 54% yield). LCMS: m/z 569 (M+H)⁺, R_(t)=0.74 min.

(i) Potassium t-butoxide (132 mg, 1.18 mmol) was added to a solution of3-(6-amino-2,3-dichloropyridin-4-yl)-6-43S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-1-benzyl-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one(67 mg, 0.12 mmol) in DMSO (3 mL). The reaction was sparged with oxygenfor 2 min, then maintained at room temperature with stirring. After 5min, the reaction was complete by LCMS analysis. At this time, thereaction was sparged with nitrogen, then neutralized with 0.6 mL 2 N aqHCl. The reaction mixture was directly purified by prep HPLC to provide3-(6-amino-2,3-dichloropyridin-4-yl)-6-43S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one(8.4 mg, 15% yield). LCMS: m/z 479 (M+H)⁺, R_(t)=0.69 min; ¹H NMR (400MHz, DMSO-d₆) δ ppm 9.56 (s, 1H), 7.76 (br s, 2H), 4.03-4.10 (m, 1H),3.66 (d, J=8.59 Hz, 1H), 3.49 (d, J=8.59 Hz, 1H), 3.43-3.47 (m, 3H),2.97-3.19 (m, 2H), 2.92 (d, J=5.05 Hz, 1H), 1.68-1.98 (m, 3H), 1.42-1.68(m, 3H), 1.08 (d, J=6.32 Hz, 3H). IC₅₀=0.38 μM.

Example 76-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(isopropylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one

General Synthetic Route Method D

(a) A solution of3-(2,3-dichloropyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(315 mg, 0.729 mmol) in isopyropylamine (3 mL) was microwave heated to150° C. for 10 min. At completion, the reaction was concentrated, thenpurified by flash chromatography [0-10% MeOH/DCM eluent] to provide3-(3-chloro-2-(isopropylamino)pyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(220 mg, 66% yield).

Steps d-e of General Synthetic Method A were applied to provide6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(isopropylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one.LCMS: m/z 487 (M+H)⁺, R_(t)=0.60 min; ¹H NMR (400 MHz, METHANOL-d₄) δppm 7.99 (d, J=5.31 Hz, 1H), 6.66 (d, J=5.05 Hz, 1H), 4.22-4.30 (m, 2H),3.84-3.93 (m, 1H), 3.73-3.84 (m, 1H), 3.46-3.55 (m, 5H), 3.25 (br d,J=4.80 Hz, 1H), 2.95-3.15 (m, 2H), 1.88-2.13 (m, 2H), 1.79-1.88 (m, 1H),1.67-1.79 (m, 1H), 1.23-1.32 (m, 9H). IC₅₀=0.034 μM.

Example 86-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(isopropylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one

General Synthetic Route Method E

(a) A solution of3-(2,3-dichloropyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(590 mg, 1.37 mmol) in DMSO (2 mL) was treated with(4-methoxyphenyl)methanamine (749 mg, 5.46 mmol) and the resultingsolution was microwave heated to 180° C. for 40 min. At completion, thereaction was partitioned between water and EtOAc. The organics werewashed with water and brine, then dried over MgSO₄ and concentrated. Thecrude was purified by flash chromatography (0-10% MeOH/DCM eluent) toprovide3-(3-chloro-2-((4-methoxybenzyl)amino)pyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(670 mg, 92% yield). LCMS: m/z 533, R_(t)=0.81 min.

Steps d-e of Method A were applied to3-(3-chloro-2-((4-methoxybenzyl)amino)pyridin-4-yl)-2-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dioneto provide3-(2-amino-3-chloropyridin-4-yl)-6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-2,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one.LCMS: m/z 445 (M+H)⁺, R_(t)=0.58 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm7.92 (d, J=5.05 Hz, 1H), 6.64 (d, J=5.05 Hz, 1H), 6.33 (s, 2H),4.00-4.11 (m, 1H), 3.66 (d, J=8.59 Hz, 1H), 3.49 (d, J=8.59 Hz, 1H),3.38 (s, 3H), 2.93-3.13 (m, 2H), 2.91 (d, J=5.05 Hz, 1H), 1.69-1.92 (m,2H), 1.51-1.67 (m, 2H), 1.08 (d, J=6.57 Hz, 3H). IC₅₀=0.036 μM.

Example 92-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one

(a) A mixture of5-bromo-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(300 mg, 0.742 mmol) in 9:1 THF:H₂O (3 mL) was treated with3-chloro-2-methoxy-4-(3,3,4,4-tetramethylborolan-1-yl)pyridine (300 mg,1.113 mmol), potassium phosphate (472 mg, 2.226 mmol), and[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (30.3 mg,0.037 mmol) under nitrogen was microwave heated to 90° C. for 2 h. Then,chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II)(30 mg, 0.038 mmol) was added, and the reaction was microwave heated to110° C. for 1 h and at 100° C. for 16 h. The reaction was concentratedand the resulting residue was purified by flash chromatograpy oversilica gel (0-40% EtOAc/heptane) to provide5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(150 mg, 43% yield). LCMS: m/z 466.7 (M+H)⁺, R_(t)=1.50 min.

(b) To a 0° C. solution of5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-2-(methylthio)-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(150 mg, 0.321 mmol) in EtOH (6 mL) was added Mo₇O₂₄(NH₄)₆—H₂O (119 mg,0.096 mmol) and 30% aq hydrogen peroxide (0.328 mL, 3.21 mmol) and theresulting mixture was stirred at room temperature for 16 h. Atcompletion, the reaction was quenched by addition of sat. aq Na₂S₂O₃.The resulting suspension was stirred at room temperature for 5 min thenpoured into a separation funnel containing brine. The mixture wasextracted with EtOAc (5×5 mL). The combined organic phases were driedover MgSO₄, filtered and concentrated to afford semi-pure5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-2-(methylsulfonyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(160 mg, 100% yield), which was used in the next step withoutpurification. LCMS: m/z 499.2 (M+H)⁺, R_(t)=1.36 min.

(c) A mixture of5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-2-(methylsulfonyl)-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(160 mg, 0.321 mmol) and(3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine-2HCl (98 mg, 0.401mmol) in DMSO (1 mL) and DIPEA (0.56 mL, 3.21 mmol) was heated to 110°C. for 2 h. At completion, the reaction was concentrated and purified byflash chromatography over silica gel (0-10% MeOH/DCM eluent) to providesemi-pure2-43S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(189 mg, 100% yield). LCMS: m/z 589.3 (M+H)⁺, R_(t)=1.07 min.

(d) To a solution of2-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one(189 mg, 0.32 mmol) in DCM (3 mL) was added TFA (1 mL). The reaction wasmaintained at room temperature for 1.5 h. The reaction was concentratedand the resulting residue was dissolved in MeOH (2 mL) and treated withethylenediamine (0.325 mL, 4.82 mmol). The resulting mixture was stirredat room temperature for 16 h. At completion, the reaction wasconcentrated and the resulting residue was purified by prep HPLC toprovide2-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-(3-chloro-2-methoxypyridin-4-yl)-3-methyl-3,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-oneas a white powder (7.5 mg, 5% yield). LCMS: m/z 459.3 (M+H)⁺, R_(t)=0.65min; ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.97 (d, J=5.31 Hz, 1H), 7.11(d, J=5.25 Hz, 1H), 4.90-5.05 (m, 1H), 4.68-4.81 (m, 1H), 4.59 (br s,1H), 4.20-4.27 (m, 1H), 4.00 (s, 3H), 3.85 (d, J=8.59 Hz, 1H), 3.71 (d,J=8.59 Hz, 1H), 3.47-3.56 (m, 3H), 3.32-3.41 (m, 4H), 3.26 (dt, J=3.28,1.64 Hz, 1H), 2.92-3.14 (m, 3H), 1.87-2.03 (m, 2H), 1.68-1.80 (m, 2H),1.22 (d, J=6.32 Hz, 3H), −0.02-0.01 (m, 1H). IC₅₀=0.026 μM.

Example 102-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-(3-chloro-2-methoxypyridin-4-yl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-one

(a) A mixture of tert-butyl5-bromo-2-chloro-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate(400 mg, 0.853 mmol),3-chloro-2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine(253 mg, 0.939 mmol), potassium phosphate (543 mg, 2.56 mmol), andPdCl₂(dppf)-DCM adduct (34.8 mg, 0.043 mmol) in THF:H₂O (10:1, 2.2 mL)was degassed, and the mixture was microwave heated to 90° C. for 2.5hours. The reaction mixture was transferred to a separatory funnelcontaining aq NH₄Cl (20 mL) and extracted with EtOAc (3×20 mL). Thecombined extracts were dried over MgSO₄, filtered and concentrated toafford a brown oil. The crude product was purified by flashchromatography over silica gel (0-40% EtOAc/heptane eluent) to provide2-chloro-5-(3-chloro-2-methoxypyridin-4-yl)-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidineas a yellow solid (48 mg, 13% yield). LCMS: m/z 431.1 (M+H)⁺,R_(t)=1.24; 41 NMR (400 MHz, DMSO-d₆) δ ppm 12.69 (br s, 1H), 8.01 (d,J=5.05 Hz, 1H), 7.68 (d, J=2.27 Hz, 1H), 7.25 (d, J=8.59 Hz, 2H), 7.10(d, J=5.05 Hz, 1H), 6.86 (d, J=8.59 Hz, 2H), 5.34 (s, 2H), 3.96 (s, 3H),3.75 (s, 3H).

(b) A suspension of2-chloro-5-(3-chloro-2-methoxypyridin-4-yl)-4-((4-methoxybenzyl)oxy)-7H-pyrrolo[2,3-d]pyrimidine(45 mg, 0.104 mmol) and(3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine (25.4 mg, 0.104mmol) in DIPEA (0.2 mL) and DMSO (0.1 mL) was heated to 120° C. andstirred for 18 h. The reaction mixture was concentrated, and the residuewas purified by prep HPLC to provide2-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-(3-chloro-2-methoxypyridin-4-yl)-3H-pyrrolo[2,3-d]pyrimidin-4(7H)-oneas a white solid (5.6 mg, 11% yield). HRMS: m/z calculated 445.1749(M+H)⁺, found 445.1758 (M+H)⁺; ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.97(d, J=5.31 Hz, 1H), 7.16 (s, 1H), 7.11 (d, J=5.25 Hz, 1H), 4.90-5.05 (m,1H), 4.68-4.81 (m, 1H), 4.59 (br s, 1H), 4.20-4.27 (m, 1H), 4.00 (s,3H), 3.85 (d, J=8.59 Hz, 1H), 3.71 (d, J=8.59 Hz, 1H), 3.47-3.56 (m,3H), 3.32-3.41 (m, 4H), 3.26 (dt, J=3.28, 1.64 Hz, 1H), 2.92-3.14 (m,3H), 1.87-2.03 (m, 2H), 1.68-1.80 (m, 2H), 1.22 (d, J=6.32 Hz, 3H),−0.02-0.01 (m, 1H). IC₅₀=0.026 μM.

Example 116-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)-1,5-dihydro-4H-pyrrolo[3,2-c]pyridin-4-one

(a) To a stirring solution of 4,6-dichloro-1H-pyrrolo[3,2-c]pyridine(1.00 g, 5.35 mmol) in DMF (14 mL) at 0° C. was added NaH (60% inmineral oil, 0.428 g, 10.69 mmol) portionwise within 5 min, and theresulting mixture was stirred at room temperature for 2 h. The reactionmixture was cooled to 0° C. and ((chloromethoxy)methyl)trimethylsilane(0.996 mL, 5.61 mmol) was added. The resulting mixture was stirred at 0°C. for 5 min and at room temperature for 3 h. The reaction mixture wasquenched via slow addition of sat. aq NH₄Cl (3 mL), transferred to aseparatory funnel containing sat. aq NH₄Cl (80 mL), and extracted withEt₂O (4×30 mL). The combined organic extracts were washed with 2 N aqNa₂CO₃ (2×75 mL), dried over MgSO₄, filtered, and concentrated in vacuoto afford a yellow oil that crystallized into a pale yellow solid uponstanding. The residue was purified by flash chromatography over silicagel (0-40% EtOAc/heptane eluent) to provide4,6-dichloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridineas a white solid (1.26 g, 74% yield). LCMS: m/z 317.1 (M+H)⁺,R_(t)=1.50; ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.62 (d, J=0.86 Hz, 1H),7.55 (d, J=3.42 Hz, 1H), 6.67 (dd, J=3.36, 0.79 Hz, 1H), 5.56 (s, 2H),3.46-3.56 (m, 2H), 0.83-0.92 (m, 2H), −0.10-0.03 (m, 9H).

(b) To a solution of4,6-dichloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridine(1.22 g, 3.85 mmol) and 4-methoxybenzyl alcohol (0.584 g, 4.23 mmol) in1,4-dioxane (12 mL) was added potassium tert-butoxide (1.726 g, 15.38mmol). The reaction mixture was stirred at room temperature for 1 h. Thereaction mixture was transferred to a separatory funnel containing sat.aq NH₄Cl (60 mL), and extracted with Et₂O (3×40 mL). The combinedextracts were dried over MgSO₄, filtered, and concentrated in vacuo toprovide6-chloro-4-((4-methoxybenzyl)oxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridineas a yellow oil (14% w/w dioxane, 1.8 g, 99% yield). LCMS: m/z 419.2(M+H)⁺, R_(t)=1.66 min; ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.47 (d,J=8.53 Hz, 2H), 7.08 (s, 1H), 7.04 (d, J=3.26 Hz, 1H), 6.93 (d, J=8.78Hz, 2H), 6.60 (d, J=3.26 Hz, 1H), 5.46 (s, 2H), 5.39 (s, 2H), 3.83 (s,3H), 3.39-3.50 (m, 2H), 0.82-0.94 (m, 2H), −0.08-0.01 (m, 9H).

(c) To a stirring solution of6-chloro-4-((4-methoxybenzyl)oxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridine(14% w/w dioxane, 770 mg, 1.580 mmol) in DMF (4 mL) at 0° C. was addedN-bromosuccinimide (309 mg, 1.739 mmol), and the resulting mixture wasstirred at 0° C. for 1.5 h. The crude product was diluted with Et₂O (5mL) and poured into a separatory funnel containing sat. aq NH₄Cl (100mL), then extracted with Et₂O (3×75 mL). The combined extracts weredried over MgSO₄, filtered, and concentrated in vacuo to afford3-bromo-6-chloro-4-((4-methoxybenzyl)oxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridineas a red oil (14% w/w DMF, 0.89 g, 97% yield). LCMS: m/z 497.1 (M+H)⁺,R_(t)=1.71 min; ¹H NMR (400 MHz, CHLOROFORM-d) 6 ppm 7.53 (d, J=8.59 Hz,2H), 7.07 (s, 1H), 7.05 (s, 1H), 6.93 (d, J=8.84 Hz, 2H), 5.51 (s, 2H),5.33 (s, 2H), 3.83 (s, 3H), 3.39-3.51 (m, 2H), 0.82-0.96 (m, 2H),−0.09-0.04 (m, 9H).

(d) To a mixture of (2,3-dichlorophenyl)boronic acid (0.484 g, 2.54mmol), K₃PO₄ (1.29 g, 6.09 mmol), and PdCl₂(dppf) (0.075 g, 1.02 mmol)under an atmosphere of nitrogen at room temperature was added a solutionof3-bromo-6-chloro-4-((4-methoxybenzyl)oxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridine(1.18 g, 2.03 mmol) in degassed THF (4 mL). To this mixture was addeddegassed water (0.4 mL), and the resulting mixture was heated to 90° C.in the microwave for 90 min Additional (2,3-dichlorophenyl)boronic acid(0.387 g, 2.03 mmol) and PdCl₂(dppf) (0.074 g, 0.102 mmol) were added,and the reaction mixture was heated to 90° C. in the microwave for 90min. The reaction mixture was transferred to a separatory funnelcontaining sat. aq NH₄Cl (50 mL) and extracted with Et₂O (3×50 mL). Thecombined extracts were dried over MgSO₄, filtered, and concentrated toafford a black oil. The residue was purified by flash chromatographyover silica (0-30% EtOAc/heptane eluant). Concentration of the purefractions provided6-chloro-3-(2,3-dichlorophenyl)-4-((4-methoxybenzyl)oxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridineas an orange oil (380 mg, 33% yield). LCMS: m/z 563.1 (M+H)⁺,R_(t)=1.79; 41 NMR (400 MHz, METHANOL-d₄) δ ppm 7.46 (dd, J=8.08, 1.52Hz, 1H), 7.31 (s, 1H), 7.26-7.29 (m, 1H), 7.25 (s, 1H), 7.16-7.22 (m,1H), 7.01-7.06 (m, 2H), 6.75-6.81 (m, 2H), 5.54 (s, 2H), 5.20 (s, 2H),3.77 (s, 3H), 3.52-3.63 (m, 2H), 0.82-0.94 (m, 2H), −0.09-0.04 (m, 9H).

(e) To a stirring solution of6-chloro-3-(2,3-dichlorophenyl)-4-((4-methoxybenzyl)oxy)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridine(260 mg, 0.461 mmol) in DCM (2.1 mL) at room temperature was added2,3-dichloro-5,6-dicyanobenzoquinone (209 mg, 0.922 mmol) and water(0.21 mL). The reaction mixture was stirred at room temperature for 17h. The reaction mixture was transferred to a separatory funnelcontaining 3:1 sat. NaHCO₃: water, (30 mL) and extracted with DCM (4×30mL). The organics were dried over MgSO₄, filtered, and concentrated toafford a brown solid. The residue was purified by flash chromatographyover silica (0-75% EtOAc/heptane eluant). Concentration of the purefractions provided6-chloro-3-(2,3-dichlorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridin-4(5H)-oneas a white solid (163 mg, 80% yield). LCMS: 443(M+H)⁺, R_(t)=1.42 min;41 NMR (400 MHz, METHANOL-d₄) δ ppm 7.48 (dd, J=7.96, 1.64 Hz, 1H),7.33-7.39 (m, 1H), 7.19-7.30 (m, 2H), 6.83 (s, 1H), 5.49 (s, 2H), 3.58(t, J=7.96 Hz, 2H), 0.89 (t, J=7.96 Hz, 2H), −0.04 (s, 9H).

(f) A solution of6-chloro-3-(2,3-dichlorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,2-c]pyridin-4(5H)-one(163 mg, 0.367 mmol) and tert-butyl((4-methylpiperidin-4-yl)methyl)carbamate (168 mg, 0.735 mmol) in DIPEA(0.75 mL) was stirred at 130° C. for 6 d. To this reaction mixture wasadded additional tert-butyl ((4-methylpiperidin-4-yl)methyl)carbamate(84 mg, 0.368 mmol) and DIPEA (0.5 mL), and the resulting mixture wasstirred for 1 d. The reaction mixture was concentrated, and the crudeproduct was dissolved in DCM (5 mL), then transferred to a separatoryfunnel containing sat. aq NH₄Cl (5 mL). The layers were separated, andthe aqueous phase was extracted with DCM (3×5 mL). The combined organicextracts were dried over MgSO₄, filtered, and concentrated to afford adark, viscous oil. The crude product was purified by flashchromatography over silica (0-50% EtOAc/DCM eluant) to providetert-butyl((1-(3-(2,3-dichlorophenyl)-4-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-4,5-dihydro-1H-pyrrolo[3,2-c]pyridin-6-yl)-4-methylpiperidin-4-yl)methyl)carbamateas an orange solid (142 mg, 61% yield). LCMS: m/z 635.4 (M+H)⁺,R_(t)=1.59 min; ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.45 (dd, J=7.96,1.64 Hz, 1H), 7.37 (dd, J=7.71, 1.64 Hz, 1H), 7.24 (t, J=7.83 Hz, 1H),7.07 (s, 1H), 6.03 (s, 1H), 3.59 (t, J=7.83 Hz, 2H) 5.45 (s, 2H),3.24-3.30 (m, 2H), 2.96-3.15 (m, 4H), 1.64 (ddd, J=13.33, 9.41, 3.79 Hz,2H), 1.40-1.51 (m, 11H), 0.98 (s, 3H), 0.85-0.94 (m, 2H), −0.08-0.01 (m,9H).

(g) To a solution of tert-butyl((1-(3-(2,3-dichlorophenyl)-4-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-4,5-dihydro-1H-pyrrolo[3,2-c]pyridin-6-yl)-4-methylpiperidin-4-yl)methyl)carbamate(142 mg, 0.223 mmol) in DCM (3 mL) was added TFA (1 mL, 12.98 mmol), andthe resulting mixture was stirred at room temperature for 8 h, then 40°C. for 1.5 h. The reaction mixture was concentrated, then redissolved inMeOH (3 mL). To the resulting solution was added ethylenediamine (0.25mL, 3.70 mmol) and the reaction mixture was stirred at room temperaturefor 16 h. The reaction mixture was concentrated and the resultingresidue was purified by prep HPLC to afford6-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)-1,5-dihydro-4H-pyrrolo[3,2-c]pyridin-4-one(36.9 mg, 40% yield). HRMS: m/z calculated 405.1243 (M+H)⁺, found405.1266 (M+H)⁺; ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.42 (dd, J=7.96,1.64 Hz, 1H), 7.37 (dd, J=7.83, 1.52 Hz, 1H), 7.22 (t, J=7.83 Hz, 1H),6.97 (s, 1H), 5.90 (s, 1H), 3.20-3.28 (m, 2H), 2.96-3.07 (m, 2H), 2.53(s, 2H), 1.63 (ddd, J=13.39, 9.73, 3.92 Hz, 2H), 1.49 (dt, J=13.52, 3.85Hz, 2H), 1.01 (s, 3H). IC₅₀=0.086 μM.

Example 12(3S,4S)-8-(7-(2,3-dichlorophenyl)-5H-pyrrolo[2,3-b]pyrazin-3-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine

(a) A 0° C. solution of 3-bromo-6-chloropyrazin-2-amine (5.00 g, 23.99mmol) in THF (80 mL) was degassed for 20 min under streaming nitrogen.The cooled solution was treated with TEA (6.7 mL, 48.0 mmol), CuI (0.457g, 2.399 mmol) and Pd(PPh₃)₂Cl₂ (0.505 g, 0.720 mmol). Then,ethynyltrimethylsilane (3.73 mL, 26.4 mmol) was slowly added to thestirred mixture. The cold bath was removed and reaction mixture wasstirred for 2 h at room temperature. At completion, the reaction mixturewas filtered through a pad of Celite and washed with EtOAc (2×100 mL).The solids were dissolved in water (100 mL) and the aqueous layer wasextracted with EtOAc (2×100 mL). The combined organic layers were driedover Na₂SO₄, filtered and concentrated. The crude was purified by flashchromatography over silica gel (0-30% EtOAc/heptane eluent) to afford6-chloro-3-((trimethylsilyl)ethynyl)pyrazin-2-amine as light brown solid(4.2 g, 78% yield). LCMS: m/z 226 (M+H)⁺, R_(t)=1.17 min.

(b) A solution of 6-chloro-3-((trimethylsilyl)ethynyl)pyrazin-2-amine(9.06 g, 40.1 mmol) in THF (150 mL) was cooled to 0° C. and treated witha suspension of t-BuOK (9.01 g, 80 mmol) in THF (50 mL). The reactionmixture was stirred at 0° C. for 30 min, then heated to reflux at 75° C.for 2.5 h after which point LCMS analysis indicated complete consumptionof the starting material. The reaction mixture was cooled to roomtemperature, then quenched with water (10 mL) and concentrated. Theresidue was partitioned between EtOAc (200 mL) and water (150 mL), andthen filtered. The organic layer was washed with water (50 mL). Thecombined aqueous layers were extracted with EtOAc (200 mL, then 100 mL).The combined organic layers were dried over Na₂SO₄, filtered andconcentrated. The crude was purified by flash chromatography over silicagel (0-30% EtOAc/heptane eluent) to afford3-chloro-5H-pyrrolo[2,3-b]pyrazine as yellow solid (2.17 g, 35% yield).LCMS: 154 (M+H)⁺, R_(t)=0.60 min.

(c) To a solution of 3-chloro-5H-pyrrolo[2,3-b]pyrazine (1.00 g, 6.51mmol) in DMF (33 mL) was added N-bromosuccinimide (1.159 g, 6.51 mmol).The reaction mixture was stirred at room temperature for 18 h. Atcompletion, the reaction was concentrated and the resulting residue wastreated with water (30 mL). The resulting slurry was stirred for 30 min,then filtered and washed with water (20 mL). The precipitate was driedfor 3 h under vacuum at 70° C. to afford crude7-bromo-3-chloro-5H-pyrrolo[2,3-b]pyrazine as brown solid (1.136 g, 48%yield). LCMS: m/z 232 (M+H)⁺, R_(t)=0.80 min.

(d) To a solution of 7-bromo-3-chloro-5H-pyrrolo[2,3-b]pyrazine (1.136g, 4.89 mmol) in DMF (25 mL) under nitrogen was added NaH (0.235 g, 5.86mmol). The resulting reaction mixture was stirred until gas evolutionsubsided. Then, 1-(chloromethyl)-4-methoxybenzene (0.795 mL, 5.86 mmol)and tetrabutlyammoniumiodide (20 mg) were added and the reaction mixturestirred for 1 h at 60° C. At completion, the reaction mixture waspartitioned between sat. aq NaHCO₃ (50 mL) and EtOAc (100 mL). Thelayers were separated and the organic layer was washed with brine (50mL). The combined aqueous layers were extracted with EtOAc (50 mL). Thecombined organic layers were dried over Na₂SO₄, then filtered andconcentrated. Residual water persisted in the crude residue. The crudewet residue was partitioned between DCM (50 mL) and water (50 mL). Thelayers were separated and the aqueous layer was extracted with DCM (50mL). The combined organic layers were dried over Na₂SO₄, then filteredand concentrated. The crude was purified by flash chromatography oversilica gel (0-40% EtOAc/heptane eluent) to afford7-bromo-3-chloro-5-(4-methoxybenzyl)-5H-pyrrolo[2,3-b]pyrazine as brightyellow solid (853 mg, 50% yield). LCMS: m/z 352 (M+H)⁺, R_(t)=1.29 min.

(e) A stirred suspension of7-bromo-3-chloro-5-(4-methoxybenzyl)-5H-pyrrolo[2,3-b]pyrazine (300 mg,0.851 mmol), (2,3-dichlorophenyl)boronic acid (162 mg, 0.851 mmol),PdCl₂(dPPf)-CH₂Cl₂ (69.5 mg, 0.085 mmol) and K₂CO₃ (470 mg, 3.40 mmol)in THF (7.1 mL) and water (1.4 mL) was degassed for 5 min then heated to50° C. for 1.5 h. LCMS analysis indicated 40% conversion of startingmaterial to desired product. The reaction mixture was partitionedbetween EtOAc (100 mL) and water (50 mL). The layers were separated andthe organic layer was washed with brine (50 mL). The organics were driedover Na₂SO₄, then filtered and concentrated. The crude was purified byflash chromatography over silica gel (0-30% EtOAc/heptane eluent) toafford3-chloro-7-(2,3-dichlorophenyl)-5-(4-methoxybenzyl)-5H-pyrrolo[2,3-b]pyrazineas a white solid (110 mg, 31% yield). LCMS: m/z 418 (M+H)⁺, R_(t)=1.58min.

(f) A solution of3-chloro-7-(2,3-dichlorophenyl)-5-(4-methoxybenzyl)-5H-pyrrolo[2,3-b]pyrazine(110 mg, 0.263 mmol) and(3S,4S)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine di-hydrochloridesalt (70.3 mg, 0.289 mmol) in DIPEA (0.525 mL) and DMSO (0.525 mL) wasstirred at 100° C. for 20 h, then 130° C. for 2.5 h. LCMS analysisindicated 30% conversion. The reaction mixture was partiallyconcentrated, then partitioned between EtOAc (50 mL) and water (25 mL).The layers were separated and the organic layer was washed with water(25 mL). The combined aqueous layers were extracted with EtOAc (25 mL).The combined organic layers were dried over Na₂SO₄, then filtered andconcentrated. The crude was purified by flash chromatography over silicagel (0-20% MeOH/DCM eluent) to afford(3S,4S)-8-(7-(2,3-dichlorophenyl)-5-(4-methoxybenzyl)-5H-pyrrolo[2,3-b]pyrazin-3-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amineas a yellow oil (36 mg, 25% yield). LCMS: m/z 552 (M+H)⁺, R_(t)=1.07min.

(g) A solution of(3S,4S)-8-(7-(2,3-dichlorophenyl)-5-(4-methoxybenzyl)-5H-pyrrolo[2,3-b]pyrazin-3-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine(36 mg, 0.065 mmol) in triflic acid (2 mL) was stirred for 2 h at 85° C.The reaction mixture was cooled in an ice bath, diluted with DCM (2 mL),then MeOH (5 mL) was slowly added. The mixture was subsequentlyconcentrated to a volume of 2 mL. This solution was diluted with MeOH (5mL), then neutralized with 7 N NH₃ in MeOH and concentrated. Theresulting crude residue was suspended in DCM (10 mL), then sonicated for5 min and filtered. The solid was purified by prep HPLC, and the purefractions were lyophilized to provide(3S,4S)-8-(7-(2,3-dichlorophenyl)-5H-pyrrolo[2,3-b]pyrazin-3-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine(2.5 mg, 8% yield). LCMS: m/z 432 (M+H)⁺, R_(t)=0.90 min; ¹H NMR (400MHz, METHANOL-d₄) δ 8.09 (s, 1H), 7.71 (d, J=7.8 Hz, 1H), 7.59 (s, 1H),7.48 (d, J=6.5 Hz, 1H), 7.33 (t, J=7.9 Hz, 1H), 4.29-4.22 (m, 1H),4.14-4.03 (m, 2H), 3.88 (d, J=8.8 Hz, 1H), 3.74 (d, J=8.8 Hz, 1H), 3.48(s, 1H), 3.15-3.11 (m, 1H), 3.06-3.01 (m, 1H), 1.87-1.68 (m, 4H), 1.23(d, 3H). IC₅₀=0.039 μM.

Example 133-((2-Amino-3-chloropyridin-4-yl)thio)-6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one

General Synthetic Route Method F

(a)7-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(1.84 g, 6.43 mmol), prepared by the method in example 6, was suspendedin acetone (32 mL). The suspension was treated with benzyl bromide (1.5mL, 12.9 mmol) and potassium carbonate (2.66 g, 19.3 mmol). Theresulting mixture was stirred at room temperature for 12 h. Water wasadded and the precipitate was filtered, washed with water and driedunder vacuum to provide2-benzyl-7-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dioneas an off-white solid (2.2 g, 90% yield). LCMS: m/z 377 (M+H)⁺,R_(t)=1.43 min. 41 NMR (400 MHz, DMSO-d₆) δ 8.63 (s, 1H), 7.41-7.25 (m,7H), 6.81 (d, J=8.6 Hz, 2H), 5.38 (s, 2H), 4.98 (s, 2H), 3.70 (s, 3H),3.20 (s, 3H).

(b) To a solution of2-benzyl-7-(4-methoxybenzyl)-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(2.18 g, 5.79 mmol) in DCM (29 mL) was added trifluoroacetic acid (4.5mL, 57.9 mmol) and trifluoromethanesulfonic acid (0.51 mL, 5.79 mmol).The resulting mixture was stirred at 23° C. for 15 min. The volatileswere removed under vacuum and a solution of sat. aq Na₂CO₃ was added.The aqueous mixture was extracted with EtOAc. The organic layers werecombined, washed with brine, dried with Na₂SO₄, then filtered andconcentrated to provide2-benzyl-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(1.4 g, 94% yield). LCMS: m/z 257 (M+H)⁺, R_(t)=0.68 min.

(c) Step d of General Method A was applied to2-benzyl-5-methyl-2,7-dihydro-4H-pyrazolo[3,4-d]pyrimidine-4,6(5H)-dione(0.65 g, 1.50 mmol) to provide6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-2-benzyl-5-methyl-2,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one(0.25 g, 71% yield). LCMS: m/z 409 (M+H)⁺, R_(t)=0.66 min.

(d) A solution of6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-2-benzyl-5-methyl-2,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one(0.36 g, 0.635 mmol) in DCM (3.2 mL) was treated with DIPEA (0.2 mL,1.27 mmol) and di-tert-butyl dicarbonate (0.21 g, 0.952 mmol). Theresulting mixture was stirred at room temperature for 10 h. Atcompletion, sat. aq NaHCO₃ was added to the reaction. The layers wereseparated and the aqueous layer was extracted with DCM. The organiclayers were combined, washed with brine, dried with Na₂SO₄, filtered andconcentrated. The crude product was purified by flash chromatographyover silica gel (10-80% EtOAc/heptane eluent) to provide tert-butyl((3S,4S)-8-(2-benzyl-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate(0.15 g, 58% yield). LCMS: m/z 509 (M+H)⁺, R_(t)=1.34 min.

(e) To a degassed solution of((3S,4S)-8-(2-benzyl-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate(0.28 g, 0.482 mmol) in MeOH (2.4 mL), was added 10% Pd—C (51 mg, 0.482mmol). The mixture was sparged with hydrogen, then stirred at roomtemperature under an atmosphere of hydrogen for 10 h. The reactionmixture was sparged with nitrogen, then filtered through Celite andconcentrated to provide tert-butyl((3S,4S)-3-methyl-8-(5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate(0.14 g, 70% yield). LCMS: m/z 419 (M+H)⁺, R_(t)=0.81 min.

(f) To a solution of tert-butyl((3S,4S)-3-methyl-8-(5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate(0.14 g, 0.34 mmol) in DMF (1.7 mL), was added N-iodosuccinimide (0.11g, 0.505 mmol). The resulting mixture was stirred at 80° C. for 2 h. Atcompletion, 20% aq Na₂S₂O₃ was added and the mixture was extracted withEtOAc. The organic layers were combined, washed with brine, dried withNa₂SO₄, then filtered and concentrated. The crude product was purifiedby flash chromatography over silica gel (0-70% EtOAc/heptane eluent) toprovide tert-butyl43S,4S)-8-(3-iodo-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate(0.17 g, 91% yield). LCMS: m/z 545 (M+H)⁺, R_(t)=0.92 min.

(g) To a solution of tert-butyl((3S,4S)-8-(3-iodo-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,4-d]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate0.17 g, 0.305 mmol) in DCM (2 mL), was added TEA (85 μL, 0.610 mmol) andacetyl chloride (32.5 μL, 0.46 mmol). The resulting mixture was stirredat room temperature for 2 h. At completion, sat. aq NaHCO₃ was added andthe mixture was extracted with EtOAc. The organic layers were combined,washed with brine, dried with Na₂SO₄, filtered and concentrated. Thecrude product was purified by flash chromatography over silica gel(10-70% EtOAc/heptane eluent) to provide tert-butyl((3S,4S)-8-(2-acetyl-3-iodo-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,44]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamateand tert-butyl((3S,4S)-8-(1-acetyl-3-iodo-5-methyl-4-oxo-4,5-dihydro-1H-pyrazolo[3,44]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamateas a mixture of regioisomers (0.138 g, 78% yield). LCMS: m/z 587 (M+H)⁺,R_(t)=1.34 min and m/z 587 (M+H)⁺, R_(t)=1.38 min.

(h) A mixture of tert-butyl((3S,4S)-8-(2-acetyl-3-iodo-5-methyl-4-oxo-4,5-dihydro-2H-pyrazolo[3,44]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamateand tert-butyl((3S,4S)-8-(1-acetyl-3-iodo-5-methyl-4-oxo-4,5-dihydro-1H-pyrazolo[3,44]pyrimidin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate(0.138 g, 0.235 mmol) in dioxane (1.2 mL) was sparged with nitrogen. Thesolution was treated with sodium 2-amino-3-chloropyridine-4-thiolate (65mg, 0.353 mmol), K₃PO₄ (0.150 g, 0.706 mmol), TMEDA (14 μL, 0.094 mmol)and CuI (9 mg, 0.047 mmol). The resulting mixture was stirred for 10 hat 100° C. At completion, the reaction mixture was filtered throughCelite. The filter cake was washed with water and the resulting aqueousfraction was washed with EtOAc. The organic layers were combined, washedwith brine, dried over Na₂SO₄, then filtered and concentrated. The crudewas dissolved in DCM (2 mL) and treated with TFA (91 μL, 1.18 mmol). Theresulting mixture was stirred at room temperature for 2 h. Atcompletion, the reaction was concentrated and the resulting residue wasazeotroped with toluene. The crude was purified by prep HPLC to provide3-((2-amino-3-chloropyridin-4-yl)thio)-6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,44]pyrimidin-4-one(3 mg, 2.4% yield). LCMS: m/z 477 (M+H)⁺, R_(t)=0.72. NMR (400 MHz,METHANOL-d₄) δ 7.55 (d, J=5.6 Hz, 1H), 6.02 (d, J=5.6 Hz, 2H), 4.37-4.16(m, 1H), 3.84 (d, J=8.7 Hz, 1H), 3.70 (d, J=8.7 Hz, 1H), 3.48 (s, 3H),3.20-3.06 (m, 2H), 3.03 (d, J=5.0 Hz, 2H), 2.08-1.85 (m, 2H), 1.83-1.65(m, 2H), 1.22 (d, J=6.5 Hz, 3H). IC₅₀=0.027 μM.

The following compounds of Table 1 were prepared according to the aboveexamples using suitable starting materials:

TABLE 1 LCMS; R_(t) (min) SHP2 IC₅₀ Example Structure Method & ¹H NMR μM14

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichlorophenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 464;R_(t) = 1.05 min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 7.72 (dd, J = 7.0,2.65 Hz, 1 H), 7.38-7.47 (m, 2 H), 4.02-4.16 (m, 1 H), 3.70 (d, J = 8.8Hz, 1 H), 3.53 (d, J = 8.6 Hz, 1 H), 2.86-3.14 (m, 3 H), 1.49-1.96 (m, 2H), 1.11 (d, J = 6.3 Hz, 3 H) 0.028 15

  6-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 421; R_(t) = 0.95 min ¹H NMR(400 MHz, METHANOL-d₄) δ ppm 7.62 (dd, J = 8.0, 1.6 Hz, 1 H), 7.40- 7.44(m, 1 H), 7.33-7.39 (m, 1 H), 3.48 (s, 3 H), 3.37 (dt, J = 13.3, 4.5 Hz,2 H), 3.19 (ddd, J = 13.1, 10.0, 2.9 Hz, 2 H), 2.59 (s, 2 H), 1.68 (ddd,J = 13.5, 9.8, 3.8 Hz, 2 H), 1.47- 1.56 (m, 2 H), 1.06 (s, 3 H) 0.034 16

  (R)-3-(2-amino-3-chloropyridin-4-yl)-6-(1-amino-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 429; R_(t) = 0.74min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 7.92 (d, J = 5.1 Hz, 1 H), 6.64 (d,J = 5.1 Hz, 1 H), 6.33 (s, 2 H), 3.38 (s, 4 H), 2.87-3.01 (m, 2 H),2.75- 2.81 (m, 1 H), 1.82-1.95 (m, 1 H), 1.19-1.80 (m, 9 H) 0.36  17

  (S)-6-(1-amino-7-azaspiro[3.5]nonan-7-yl)-3-(2,3-dichlorophenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 433; R_(t) = 1.29 min ¹H NMR(400 MHz, METHANOL-d₄) δ ppm 7.62 (dd, J = 7.8, 1.8 Hz, 1 H), 7.40- 7.45(m, 1 H), 7.33-7.39 (m, 1 H), 3.39-3.61 (m, 5 H), 3.01- 3.17 (m, 2 H),2.86-2.99 (m, 1 H), 2.18-2.30 (m, 1 H), 1.71- 2.00 (m, 5 H), 1.49-1.68(m, 2 H) 0.044 18

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-methoxypyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 460; R_(t) =0.48 min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.01 (d, J = 5.1 Hz, 1 H),6.57-6.64 (m, 2 H), 4.06 (dd, J = 6.3, 5.05 Hz, 1 H), 3.66 (d, J = 8.6Hz, 1 H), 3.49 (d, J = 8.6 Hz, 1 H), 3.37 (s, 3 H), 2.94-3.11 (m, 2 H),2.86- 2.93 (m, 4 H), 1.68-1.92 (m, 2 H), 1.51-1.65 (m, 2 H), 1.08 (d, J= 6.3 Hz, 3 H) 0.043 19

  (R)-6-(1-amino-8-azaspiro[4.5]decan-8-yl)-5-methyl-3-phenyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one B(M + H)⁺ = 379; R_(t) = 0.76 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm8.13- 8.23 (m, 2 H), 7.31-7.48 (m, 3 H), 3.40-3.61 (m, 5 H), 2.96- 3.10(m, 2 H), 2.85-2.92 (m, 1 H), 2.77-2.89 (m, 1 H), 1.96- 2.11 (m, 1 H),1.32-1.94 (m, 10 H) 0.046 20

  (S)-6-(4-amino-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichlorophenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 449; R_(t) = 1.73min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.62 (dd, J = 7.8, 1.8 Hz, 1 H),7.40- 7.44 (m, 1 H), 7.37 (d, J = 7.8 Hz, 1 H), 4.12 (dd, J = 9.1, 6.6Hz, 1 H), 3.74-3.87 (m, 2 H), 3.44-3.57 (m, 6 H), 3.18 (t, J = 5.8 Hz, 1H), 2.99-3.12 (m, 2 H), 1.83-1.98 (m, 2 H), 1.59- 1.71 (m, 2 H) 0.050 21

  (S)-6-(4-amino-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloropyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 450; R_(t) = 0.88min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.37 (d, J = 5.1 Hz, 1 H), 7.53(d, J = 4.8 Hz, 1 H), 4.13 (dd, J = 9.0, 6.4 Hz, 1 H), 3.74-3.87 (m, 2H), 3.42-3.59 (m, 6 H), 3.18 (t, J = 5.8 Hz, 1 H), 2.98- 3.13 (m, 2 H),1.82-1.99 (m, 2 H), 1.58-1.78 (m, 2 H) 0.054 22

  (R)-6-(1-amino-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloropyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 448; R_(t) =0.89 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.37 (d, J = 4.8 Hz, 1 H),7.53 (d, J = 4.8 Hz, 1 H), 3.42-3.65 (m, 5 H), 2.98-3.17 (m, 2 H), 2.86(t, J = 7.3 Hz, 1 H), 1.98-2.20 (m, 1 H), 1.25-1.93 (m, 9 H) 0.057 23

  (S)-3-(2-amino-3-chloropyridin-4-yl)-6-(4-amino-2-oxa-8-azaspiro[4.5]decan- 8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 431; R_(t) = 0.45 min ¹H NMR(400 MHz, DMSO- d₆) δ ppm 7.92 (d, J = 5.1 Hz, 1 H), 6.64 (d, J = 4.8Hz, 1 H), 6.33 (s, 2 H), 3.95 (t, J = 7.5 Hz, 1 H), 3.54-3.73 (m, 2 H),3.38 (s, 4 H), 3.07 (br. s., 1 H), 2.89- 3.00 (m, 2 H), 1.64-1.88 (m, 2H), 1.49 (br. s., 2 H) 0.064 24

  6-(7-amino-3-azaspiro[5.5]undecan-3-yl)-3-(2,3-dichlorophenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 461; R_(t) = 0.87 min ¹HNMR (400 MHz, DMSO- d₆) δ ppm 7.72 (dd, J = 7.3, 2.5 Hz, 1 H), 7.39-7.47(m, 2 H), 3.26-3.35 (m, 2 H), 2.94- 3.10 (m, 2 H), 2.46 (d, J = 4.5 Hz,1 H), 1.87-2.06 (m, 2 H), 1.67-1.78 (m, 1 H), 1.21- 1.65 (m, 9 H), 1.13(d, J = 14.9 Hz, 1 H), 0.99-1.08 (m, 1 H) 0.072 25

  6-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-(2,3-dichloropyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 422; R_(t) = 0.73 min ¹HNMR (400 MHz, METHANOL-d₄) δ ppm 8.37 (d, J = 4.8 Hz, 1 H), 7.53 (d, J =5.1 Hz, 1 H), 3.49 (s, 3 H), 3.35-3.44 (m, 2 H), 3.15- 3.26 (m, 2 H),2.60 (s, 2 H), 1.63-1.75 (m, 2 H), 1.52 (d, J = 14.1 Hz, 2 H), 1.06 (s,3 H). 0.10  26

  6-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-(2-chloro-3-(trifluoromethyl)phenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 455; R_(t) =1.40 ¹H NMR (400 MHz, DMSO- d₆) δ ppm 7.94 (dd, J = 7.8, 1.3 Hz, 1 H),7.75-7.80 (m, 1 H), 7.58-7.66 (m, 1 H), 3.36 (s, 3 H), 3.21-3.30 (m, 2H), 3.02- 3.13 (m, 2 H), 2.43 (s, 2 H), 1.52-1.63 (m, 2 H), 1.31- 1.42(m, 2 H), 0.93 (s, 3 H) 0.127 27

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dichloro-6-fluorophenyl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 481; R_(t) =1.05 ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.67 (dd, J = 9.1, 5.31 Hz, 1H), 7.23 (t, J = 8.7 Hz, 1 H), 4.19-4.27 (m, 1 H), 3.85 (d, J = 8.6 Hz,1 H), 3.71 (d, J = 8.84 Hz, 1 H), 3.40-3.58 (m, 6 H), 3.04 (d, J = 5.1Hz, 2 H), 1.70-1.80 (m, 1 H), 1.22 (d, J = 6.6 Hz, 4 H) 0.45  28

  6-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-(3-chloropyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one B (M + H)⁺ = 388; R_(t) = 0.81 min ¹HNMR (400 MHz, METHANOL-d₄) δ ppm 8.68 (s, 1 H), 8.47-8.58 (m, 1 H), 7.62(d, J = 5.1 Hz, 1 H), 3.49 (s, 3 H), 3.34-3.44 (m, 2 H), 3.15-3.25 (m, 2H), 2.60 (s, 2 H), 1.68 (ddd, J = 13.4, 9.9, 3.8 Hz, 2 H), 1.41-1.58 (m,3 H), 1.03-1.14 (m, 3 H) 0.312 29

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(cyclobutylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 499; R_(t) = 0.62min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.97 (d, J = 4.9 Hz, 1 H), 6.68(d, J = 5.1 Hz, 1 H), 4.52 (quin, J = 8.15 Hz, 1 H), 4.19-4.26 (m, 1 H),3.82-3.97 (m, 1 H), 3.81-3.88 (m, 1 H), 3.66- 3.72 (m, 1 H), 3.62-3.77(m, 1 H), 3.38-3.53 (m, 5 H), 2.96- 3.19 (m, 3 H), 2.31-2.47 (m, 2 H),1.86-2.10 (m, 4 H), 1.66- 1.84 (m, 4 H), 1.22 (d, J = 6.6 Hz, 3 H) 0.03530

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-morpholinopyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 515; R_(t) = 0.53min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.26 (d, J = 5.1 Hz, 1 H), 7.11 (d,J = 4.8 Hz, 1 H), 4.02-4.11 (m, 1 H), 3.71- 3.82 (m, 4 H), 3.61-3.71 (m,1 H), 3.49 (d, J = 8.3 Hz, 2 H), 3.36-3.40 (m, 4 H), 2.89- 3.17 (m, 4H), 1.80-1.98 (m, 1 H), 1.68-1.80 (m, 2 H), 1.42- 1.68 (m, 3 H), 1.08(d, J = 6.3 Hz, 3 H) 0.103 31

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(pyrrolidin-1-yl)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 499; R_(t) = 1.03min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.07 (d, J = 4.8 Hz, 1 H), 6.76 (d,J = 4.8 Hz, 1 H), 4.02-4.10 (m, 1 H), 3.66 (d, J = 8.6 Hz, 1 H),3.55-3.63 (m, 4 H), 3.49 (d, J = 8.3 Hz, 1 H), 3.38 (s, 3 H), 2.93-3.12(m, 2 H), 2.91 (d, J = 5.1 Hz, 1 H), 1.81-1.97 (m, 5 H), 1.69- 1.81 (m,1 H), 1.50-1.65 (m, 2 H), 1.08 (d, J = 6.3 Hz, 3 H) 0.012 32

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(methylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 459; R_(t) = 1.03min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.01 (d, J = 5.1 Hz, 1 H),6.57-6.64 (m, 2 H), 4.06 (dd, J = 6.3, 5.05 Hz, 1 H), 3.66 (d, J = 8.6Hz, 1 H), 3.49 (d, J = 8.6 Hz, 1 H), 3.37 (s, 3 H), 2.94-3.11 (m, 2 H),2.86- 2.93 (m, 4 H), 1.68-1.92 (m, 2 H), 1.51-1.65 (m, 2H), 1.08 (d, J =6.3 Hz, 3 H) 0.054 33

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-bromo-2- chloropyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 508; R_(t) = 0.55min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.40 (d, J = 4.8 Hz, 1 H), 7.44(d, J = 4.8 Hz, 1 H), 4.20-4.27 (m, 1 H), 3.85 (d, J = 8.6 Hz, 1 H),3.71 (d, J = 8.8 Hz, 1 H), 3.51 (s, 3 H), 3.43-3.50 (m, 2 H), 2.96-3.21(m, 4 H), 1.85- 2.03 (m, 2 H), 1.65-1.80 (m, 2 H), 1.22 (d, J = 6.3 Hz,3 H) 0.013 34

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2-bromopyridin- 4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 474, 476; R_(t) = 0.56min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.78 (s, 1 H), 8.44 (d, J = 5.3 Hz,1 H), 8.33 (d, J = 5.1 Hz, 1 H), 4.07 (br s, 1 H), 3.66 (br s, 1 H),3.39-3.54 (m, 5 H), 2.95-3.14 (m, 3 H), 1.52- 1.98 (m, 4 H), 1.09 (br s,3 H) 0.027 35

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-bromo-2(methylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 503, 505; R_(t) =0.49 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.02 (d, J = 5.1 Hz, 1 H),6.62 (d, J = 5.1 Hz, 1 H), 4.20-4.26 (m, 1 H), 3.66-3.74 (m, 1 H), 3.85(d, J = 8.6 Hz, 1 H), 3.38-3.56 (m, 6 H), 3.06-3.17 (m, 1 H), 3.03 (d, J= 5.1 Hz, 2 H), 2.99 (s, 3 H), 2.64-2.68 (m, 1 H), 1.87-2.07 (m, 3 H),1.68- 1.87 (m, 3 H) 1.22 (d, J = 6.3 Hz, 3 H) 0.034 36

  6-((1R,3R)-1-amino-3-methyl-8- azaspiro[4.5]decan-8-yl)-5-methyl-3-(pyridin-4-yl)-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one A (M +H)⁺ = 394; R_(t) = 1.10 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.48 (d,J = 6.1 Hz, 2 H), 8.33 (d, J = 6.1 Hz, 2 H), 3.46 (s, 3 H), 3.34-3.44(m, 2 H), 2.94 (br t, J = 12.4 Hz, 2 H), 2.73-2.82 (m, 1 H), 2.61 (dd, J= 10.5, 1.64 Hz, 1 H), 2.54 (d, J = 9.4 Hz, 1 H), 2.00-2.12 (m, 1 H),1.90-2.00 (m, 1 H), 1.65- 1.90 (m, 3 H), 1.34 (br t, J = 11.1 Hz, 2 H),1.12-1.25 (m, 1 H), 1.00-1.10 (m, 1 H), 0.97 (d, J = 6.3 Hz, 3 H) 0.00937

  3-(2-amino-3-chloropyridin-4-yl)-6-((1R,3S)-1-amino-3-fluoro-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺= 447; R_(t) = 0.49 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 7.92 (d, J =5.1 Hz, 1 H), 6.76 (d, J = 5.3 Hz, 1 H), 5.20 (br t, J = 5.7 Hz, 1 H),4.99-5.30 (m, 1 H), 5.06 (br t, J = 5.7 Hz, 1 H), 3.50 (s, 5 H),3.11-3.21 (m, 1 H), 3.04 (br t, J = 12.5 Hz, 2 H), 2.13-2.36 (m, 2 H),1.73- 2.08 (m, 5 H), 1.52 (br d, J = 11.9 Hz, 1 H), 1.37 (br d, J = 12.9Hz, 1 H) 0.027 38

  6-((1R,3S)-1-amino-3-fluoro-8- azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(cyclopropylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 487; R_(t) =0.98 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.02- 8.07 (m, 1 H),6.74-6.77 (m, 1 H), 5.04-5.37 (m, 1 H), 3.45- 3.63 (m, 6 H), 2.92-3.18(m, 3 H), 2.69-2.79 (m, 1 H), 2.25 (td, J = 15.5, 6.19 Hz, 3 H), 1.87-2.10 (m, 4 H), 1.76-1.87 (m, 4H), 1.37-1.73 (m, 1 H), 1.35- 1.36 (m, 1H), 1.35-1.37 (m, 1 H), 0.76-0.89 (m, 2 H), 0.53- 0.64 (m, 2 H) 0.011 39

  6-((1R,3S)-1-amino-3-fluoro-8- azaspiro[4.5]decan-8-yl)-5-methyl-3-phenyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ =397; R_(t) = 0.63 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.17- 8.23 (m,2 H), 7.34-7.48 (m, 2 H), 5.02-5.24 (m, 1 H), 3.47- 3.59 (m, 5 H), 3.16(dd, J = 9.5, 6.69 Hz, 1 H), 2.95- 3.06 (m, 2 H) 2.14-2.31 (m, 2 H),1.75-2.01 (m, 4 H), 1.51 (br d, J = 13.4 Hz, 1 H), 1.36 (br d, J = 12.9Hz, 1 H) 0.037 40

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(6-(dimethylamino)-2-methylpyridin-3-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 453; R_(t) = 0.96 min ¹HNMR (400 MHz, METHANOL-d₄) δ ppm 7.55 (d, J = 8.8 Hz, 1 H), 6.52 (d, J =8.6 Hz, 1 H), 4.20-4.27 (m, 1 H), 3.85 (d, J = 8.6 Hz, 1 H), 3.70 (d, J= 8.84 Hz, 1 H), 3.50 (s, 3 H), 3.37-3.47 (m, 2 H), 3.11 (s, 7 H),2.98-3.08 (m, 2 H), 2.34-2.40 (m, 3 H), 1.86- 2.01 (m, 2 H), 1.62-1.86(m, 2 H) 1.22 (d, J = 6.6 Hz, 3 H) 0.089 41

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-3-phenyl-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one A (M + H)⁺ =395; R_(t) = 0.79 min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.33 (dd, J =8.5, 1.4 Hz, 1 H), 7.33-7.46 (m, 1 H), 4.02-4.10 (m, 1 H), 3.66 (d, J =8.3 Hz, 1 H), 3.49 (d, J = 8.3 Hz, 1 H), 3.43-3.46 (m, 3 H), 2.88-3.10(m, 3 H), 1.82- 1.92 (m, 1 H), 1.70-1.80 (m, 1 H), 1.52-1.66 (m, 2 H),1.09 (d, J = 6.3 Hz, 3 H) 0.033 42

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8- azaspiro[4.5]decan-8-yl)-3-(4-methoxyphenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 425; R_(t) = 0.99 min ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm8.23 (br d, J = 8.6 Hz, 2 H), 6.90 (d, J = 8.8 Hz, 2 H), 4.12 (dd, J =6.3, 4.8 Hz, 1 H), 3.76 (br s, 4 H), 3.64 (d, J = 8.8 Hz, 1 H), 3.48 (s,3 H), 3.19-3.37 (m, 2 H), 2.99 (br d, J = 3.5 Hz, 3 H), 1.85-2.02 (m, 2H), 1.63- 1.84 (m, 4 H), 1.19 (br d, J = 6.3 Hz, 3 H) 0.020 43

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-((R)-3-hydroxypyrrolidin-1-yl)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one D (M + H)⁺ =515; R_(t) = 0.47 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.04 (d, J =5.1 Hz, 1 H), 6.79 (d, J = 4.8 Hz, 1 H), 4.48 (br s, 1 H), 4.30 (br dd,J = 6.4, 4.2 Hz, 1 H), 3.90-4.00 (m, 3 H), 3.84 (d, J = 9.1 Hz, 1 H),3.65-3.75 (m, 1 H) 3.45-3.61 (m, 7 H), 2.93-3.26 (m, 5 H), 1.64- 2.15(m, 8 H), 1.32 (dd, J = 6.7, 4.2 Hz, 5 H) 0.084 44

  6-((3S,4S)-4-amino-3-methyl-2- oxa-8-azaspiro[4.5]decan-8-yl)-3-(2-chloropyridin-4-yl)-5-methyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 430; R_(t) = 0.92 min ¹H NMR (400 MHz,DMSO- d₆) δ ppm 8.64 (s, 1 H), 8.47 (d, J = 5.3 Hz, 1 H), 8.30 (dd, J =5.3, 1.26 Hz, 1 H), 4.06 (quin, J = 6.0 Hz, 1 H), 3.66 (d, J = 8.6 Hz, 1H), 3.49 (d, J = 8.6 Hz, 1 H), 2.96-3.14 (m, 2 H), 2.88-2.96 (m, 1 H),2.48- 2.53 (m, 5 H), 1.81-1.94 (m, 1 H), 1.70-1.81 (m, 1 H), 1.49- 1.68(m, 2 H), 1.09 (d, J = 6.3 Hz, 3 H) 0.016 45

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2-aminopyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 411; R_(t) = 0.65 min ¹HNMR (400 MHz, DMSO- d₆) δ ppm 7.83-8.12 (m, 1 H), 7.39-7.52 (m, 2 H),5.91 (br s, 2 H), 4.06 (br s, 1 H), 3.66 (br d, J = 8.6 Hz, 1 H),3.47-3.51 (m, 1 H), 3.49 (br d, J = 8.3 Hz, 1 H), 3.40-3.46 (m, 2 H),2.88- 3.14 (m, 3 H), 1.87 (br s, 1 H), 1.68-1.82 (m, 1 H), 1.61 (br d, J= 14.7 Hz, 2 H), 1.00-1.16 (m, 3 H) 0.037 46

  6-((3S,4S)-4-amino-3-methyl-2- oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-3-(2-(methylamino)pyridin- 4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 425; R_(t) = 0.49 min ¹H NMR (400 MHz,DMSO- d₆) δ ppm 8.01 (d, J = 5.3 Hz, 1 H), 7.58 (s, 1 H), 7.38 (dd, J =5.4, 1.1 Hz, 1 H), 6.50 (br d, J = 4.8 Hz, 1 H), 4.13-4.17 (m, 1 H),4.11-4.19 (m, 1 H), 4.03- 4.11 (m, 1 H), 3.62-3.74 (m, 1 H), 3.52 (br d,J = 8.3 Hz, 1 H), 3.42-3.47 (m, 3 H), 2.89- 3.13 (m, 3 H), 2.80 (d, J =4.8 Hz, 3 H), 1.50-1.93 (m, 5 H), 1.10 (d, J = 6.3 Hz, 3 H) 0.050 47

  3-(6-amino-2-methylpyridin-3-yl)- 6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 425; R_(t) = 0.57 min ¹H NMR (400 MHz,METHANOL-d₄) δ ppm 8.15 (d, J = 7.7 Hz, 1 H), 6.95-7.03 (m, 1 H),4.20-4.27 (m, 1 H), 3.64-3.87 (m, 4 H), 3.38- 3.50 (m, 1 H), 2.95-3.17(m, 2 H), 1.85-2.01 (m, 1 H), 1.74 (br t, J = 13.3 Hz, 1 H), 1.16- 1.28(m, 2 H) 0.037 48

  3-(6-amino-2-methylpyridin-3-yl)- 6-((1R,3R)-1-amino-3-methyl-8-azaspiro[4.5]decan-8-yl)-5-methyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 423; R_(t) = 0.72 min ¹H NMR (400 MHz,METHANOL-d₄) δ ppm 1.03- 1.10 (m, 3 H) 1.16 (dt, J = 12.4, 9.85 Hz, 1 H)1.28 (dd, J = 12.9, 9.35 Hz, 1 H) 1.37-1.52 (m, 2 H) 1.70-1.89 (m, 2 H)1.89- 2.10 (m, 2 H) 2.12-2.24 (m, 1 H) 2.92 (dd, J = 9.5, 6.44 Hz, 1 H)2.96-3.07 (m, 2 H) 3.41- 3.50 (m, 2 H) 3.53 (s, 3 H) 3.84 (s, 3 H) 6.98(d, J = 7.7 Hz, 1 H) 8.11-8.19 (m, 1 H) 0.016 49

  (R)-3-(6-amino-2-methylpyridin-3- yl)-6-(1-amino-3,3-difluoro-8-azaspiro[4.5]decan-8-yl)-5- methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 445; R_(t) = 1.11 min ¹H NMR(400 MHz, METHANOL-d₄) δ ppm 1.44- 1.64 (m, 2 H) 1.81-2.21 (m, 1 H)1.84-1.93 (m, 1 H) 1.97- 2.18 (m, 2 H) 2.41-2.58 (m, 2 H) 2.93-3.07 (m,2 H) 3.15 (br t, J = 8.5 Hz, 1 H) 3.47-3.60 (m, 5 H) 3.84 (s, 3 H) 6.98(d, J = 8.8 Hz, 1 H) 8.15 (d, J = 8.8 Hz, 1 H) 0.009 50

  (R)-6-(1-amino-3,3-difluoro-8- azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(cyclopropylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 505; R_(t) = 1.01min ¹H NMR (400 MHz, DMSO- d₆) δ ppm 8.07 (d, J = 5.1 Hz, 1 H), 6.68 (d,J = 5.0 Hz, 1 H), 6.60-6.67 (m, 1 H), 3.27- 3.55 (m, 8 H), 2.72-3.09 (m,5 H), 2.27-2.48 (m, 3 H), 1.66- 2.12 (m, 6 H), 1.27-1.45 (m, 3 H),0.62-0.81 (m, 2 H), 0.51- 0.62 (m, 2 H) 0.035 51

  (R)-6-(1-amino-3,3-difluoro-8- azaspiro[4.5]decan-8-yl)-5-methyl-3-(pyridin-4-yl)-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺= 416; R_(t) = 0.53 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.58 (br d,J = 5.3 Hz, 2 H), 8.43 (d, J = 6.3 Hz, 2 H), 3.52-3.63 (m, 5 H),2.94-3.17 (m, 3 H), 2.41- 2.55 (m, 2 H), 1.82-2.18 (m, 5 H), 1.43-1.63(m, 2 H) 0.014 52

  3-(6-amino-2-chloropyridin-3-yl)-6- ((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 445; R_(t) = 0.85 min ¹H NMR (400 MHz,METHANOL-d₄) δ ppm 7.54- 7.56 (m, 1 H), 4.19-4.27 (m, 1 H), 3.84 (d, J =8.8 Hz, 1 H), 3.63-3.77 (m, 1 H), 3.52- 3.59 (m, 3 H), 3.39-3.50 (m, 2H), 2.94-3.20 (m, 3 H), 1.84- 2.03 (m, 2 H), 1.65-1.84 (m, 2 H),1.12-1.30 (m, 3 H) 0.023 53

  3-(6-amino-2-methylpyridin-3-yl)-6-((1R,3R)-1-amino-3-(trifluoromethyl)-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4- one E (M + H)⁺ = 477; R_(t) =0.72 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm 8.13- 8.18 (m, 1 H), 6.98(d, J = 7.8 Hz, 1 H), 4.85 (s, 5 H), 3.84 (s, 1 H), 3.40-3.55 (m, 3 H),2.77- 2.94 (m, 1 H), 2.25 (ddd, J = 13.1, 8.53, 6.44 Hz, 1 H), 1.73-1.95(m, 2 H), 1.58- 1.73 (m, 1 H), 1.39-1.58 (m, 1H) 0.036 54

  6-((1R,3R)-1-amino-3-(trifluoromethyl)-8-azaspiro[4.5]decan-8-yl)-5-methyl- 3-(pyridin-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 448; R_(t) = 0.61 min ¹H NMR(400 MHz, METHANOL-d₄) δ ppm 8.58 (d, J = 6.1 Hz, 2 H), 8.43 (d, J = 5.2Hz, 2 H), 3.57 (s, 3 H), 3.46-3.56 (m, 2 H), 3.01- 3.14 (m, 2 H),2.91-3.00 (m, 1 H), 2.86 (br d, J = 8.8 Hz, 1 H), 2.19-2.28 (m, 1 H),1.82- 1.96 (m, 4 H), 1.64 (dt, J = 13.0, 9.7 Hz, 1 H), 1.40-1.55 (m, 2H) 0.021 55

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(3-aminophenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 410; R_(t) = 0.49 min ¹H NMR (400 MHz, METHANOL-d₄) δ ppm7.54 (s, 1 H), 7.50 (br d, J = 7.3 Hz, 1 H), 7.18 (t, J = 7.8 Hz, 1 H),6.78 (ddd, J = 8.0, 2.3, 0.9 Hz, 1 H), 4.25-4.32 (m, 1 H), 3.93 (d, J =9.1 Hz, 1 H), 3.82 (d, J = 9.1 Hz, 1 H), 3.45-3.58 (m, 5 H), 3.39 (d, J= 4.0 Hz, 1 H), 2.92-3.17 (m, 2 H), 1.96 (br d, J = 11.4 Hz, 2 H), 1.83-1.92 (m, 1 H), 1.74 (br d, J = 13.1 Hz, 1 H), 1.30 (d, J = 6.6 Hz, 3 H)0.083 56

  6-((3S,4S)-4-amino-3-methyl-2- oxa-8-azaspiro[4.5]decan-8-yl)-3-(2,3-dihydrobenzofuran-5-yl)-5- methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 437.2; R_(t) = 1.08 ¹H NMR(400 MHz, METHANOL d₄) δ 8.20-8.10 (m, 1 H), 8.08-7.88 (m, 1 H),7.04-6.73 (m, 1 H), 4.59 (t, J = 8.7 Hz, 1 H), 4.28-4.18 (m, 1 H),3.88-3.82 (m, 2 H), 3.70 (d, J = 8.7 Hz, 1 H), 3.54 (s, 3 H), 3.48-3.38(m, 2 H), 3.29- 3.23 (m, 2 H), 3.15-3.06 (m, 1 H), 3.06-2.96 (m, 2 H),2.01- 1.84 (m, 2 H), 1.82-1.67 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H).0.095 57

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8- azaspiro[4.5]decan-8-yl)-3-(4-fluorophenyl)-5-methyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 413.2; R_(t) = 1.62 ¹H NMR (400 MHz, METHANOL-d₄) δ 8.32-8.23(m, 2 H), 7.15 (t, J = 8.9 Hz, 2 H), 4.28-4.20 (m, 1 H), 3.85 (d, J =8.7 Hz, 1 H), 3.71 (d, J = 8.7 Hz, 1 H), 3.55 (s, 3 H), 3.50-3.41 (m, 2H), 3.17- 3.08 (m, 1 H), 3.08-2.97 (m, 2 H), 2.02-1.87 (m, 2 H), 1.82-1.68 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H). 0.011 58

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8- azaspiro[4.5]decan-8-yl)-3-(3,4-difluorophenyl)-5-methyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-oneA (M + H)⁺ = 431.2; R_(t) = 1.85 ¹H NMR (400 MHz, METHANOL-d₄) δ8.41-8.31 (m, 1 H), 8.17-8.09 (m, 1 H), 7.36-7.25 (m, 1 H), 4.28- 4.19(m, 1 H), 3.85 (d, J = 8.7 Hz, 1 H), 3.70 (d, J = 8.7 Hz, 1 H), 3.56 (s,3 H), 3.45 (dd, J = 13.7, 4.7 Hz, 2 H), 3.17- 3.08 (m, 1 H), 3.08-2.97(m, 2 H), 2.01-1.86 (m, 2 H), 1.81- 1.67 (m, 2 H), 1.22 (d, J = 6.5 Hz,3 H). 0.011 59

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-fluoro-4-methoxyphenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 443.2; R_(t) = 1.66 ¹H NMR (400 MHz, METHANOL-d₄) δ 8.12 (d,J = 13.1, 2.1 Hz, 1 H), 8.07 (d, J = 8.7 Hz, 1 H), 7.14 (t, J = 8.7 Hz,1 H), 4.28-4.19 (m, 1 H), 3.92 (s, 3 H), 3.85 (d, J = 8.7 Hz, 1 H), 3.70(d, J = 8.7 Hz, 1 H), 3.55 (s, 3 H), 3.51-3.39 (m, 2 H), 3.17-3.08 (m, 1H), 3.08-2.98 (m, 2 H), 2.01- 1.87 (m, 2 H), 1.82-1.68 (m, 2 H), 1.22(d, J = 6.5 Hz, 3 H). 0.013 60

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2,3-difluorophenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-oneA (M + H)⁺ = 431.2; R_(t) = 1.01 ¹H NMR (400 MHz, METHANOL-d₄) δ 7.51(t, J = 6.9 Hz, 1 H), 7.38-7.29 (m, 1 H), 7.28-7.17 (m, 1 H), 4.28- 4.19(m, 1 H), 3.85 (d, J = 8.7 Hz, 1 H), 3.71 (d, J = 8.7 Hz, 1 H), 3.52 (s,3 H), 3.51-3.42 (m, 2 H), 3.19-3.10 (m, 1 H), 3.10-3.00 (m, 2 H), 2.02-1.87 (m, 2 H), 1.82-1.69 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H). 0.013 61

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(3-fluorophenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 413.2; R_(t) = 1.70 ¹H NMR (400 MHz, METHANOL-d₄) δ 8.16-8.11(m, 1 H), 8.09 (d, J = 7.9 Hz, 1 H), 7.48-7.36 (m, 1 H), 7.17- 7.04 (m,1 H), 4.27-4.19 (m, 1 H), 3.85 (d, J = 8.7 Hz, 1 H), 3.71 (d, J = 8.7Hz, 1 H), 3.56 (s, 3 H), 3.51-3.41 (m, 2 H), 3.17-3.08 (m, 1 H), 3.08-2.99 (m, 2 H), 2.02-1.86 (m, 2 H), 1.82-1.67 (m, 2 H), 1.22 (d, J = 6.5Hz, 3 H). 0.014 62

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2,5-difluorophenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-oneA (M + H)⁺ = 431.2; R_(t) = 1.01 ¹H NMR (400 MHz, METHANOL-d₄) δ 7.55(s, 1 H), 7.28-7.13 (m, 2 H), 4.30- 4.16 (m, 1 H), 3.85 (d, J = 8.7 Hz,1 H), 3.71 (d, J = 8.7 Hz, 1 H), 3.53 (s, 3 H), 3.50-3.42 (m, 2 H),3.18-3.10 (m, 1 H), 3.09-3.01 (m, 2 H), 2.01- 1.87 (m, 2 H), 1.81-1.68(m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H). 0.017 63

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2-fluorophenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 413.2; R_(t) = 1.31 ¹H NMR (400 MHz, METHANOL-d₄) δ 7.71 (t,J = 6.7 Hz, 1 H), 7.50-7.42 (m, 1 H), 7.26 (t, J = 7.6, 1.1 Hz, 1 H),7.24-7.17 (m, 1 H), 4.27- 4.20 (m, 1 H), 3.85 (d, J = 8.7 Hz, 1 H), 3.71(d, J = 8.7 Hz, 1 H), 3.52 (s, 3 H), 3.50-3.41 (m, 2 H), 3.17-3.09 (m, 1H), 3.09-3.00 (m, 2 H), 2.02- 1.86 (m, 2 H), 1.83-1.68 (m, 3 H), 1.22(d, J = 6.5 Hz, 3 H). 0.023 64

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8- azaspiro[4.5]decan-8-yl)-3-(6-methoxypyridin-3-yl)-5-methyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 426.2; R_(t) = 1.98 ¹H NMR (400 MHz, METHANOL-d₄) δ9.06 (s, 1 H), 8.50 (d, J = 8.7, 2.4 Hz, 1 H), 6.87 (d, J = 9.3 Hz, 1H), 4.28-4.20 (m, 1 H), 3.96 (s, 3 H), 3.85 (d, J = 8.7 Hz, 1 H), 3.71(d, J = 8.7 Hz, 1 H), 3.56 (s, 3 H), 3.50-3.42 (m, 2 H), 3.16-3.09 (m, 1H), 3.09- 2.99 (m, 2 H), 2.02-1.88 (m, 2 H), 1.82-1.68 (m, 2 H), 1.22(d, J = 6.4 Hz, 3 H). 0.024 65

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(3,5-difluorophenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-oneA (M + H)⁺ = 431.2; R_(t) = 1.95 ¹H NMR (400 MHz, METHANOL-d₄) δ8.11-8.01 (m, 2 H), 6.93 (tt, J = 9.0, 2.4 Hz, 1 H), 4.28-4.17 (m, 1 H),3.85 (d, J = 8.7 Hz, 1 H), 3.70 (d, J = 8.7 Hz, 1 H), 3.56 (s, 3 H),3.52-3.40 (m, 2 H), 3.17- 3.09 (m, 1 H), 3.09-2.98 (m, 2 H), 2.02-1.85(m, 2 H), 1.81- 1.67 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H). 0.038 66

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2-methoxyphenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 425.2; R_(t) = 1.37 ¹H NMR (400 MHz, METHANOL-d₄) δ 7.54-7.30(m, 2 H), 7.16-7.07 (m, 1 H), 7.07-6.96 (m, 1 H), 4.28- 4.20 (m, 1 H),3.92-3.75 (m, 4 H), 3.71 (d, J = 8.7 Hz, 1 H), 3.51 (s, 3 H), 3.41 (s, 2H), 3.09-2.93 (m, 3 H), 2.00- 1.85 (m, 2 H), 1.80-1.71 (m, 2 H), 1.22(d, J = 6.5 Hz, 3 H). 0.091 67

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl-3- (2-methylpyridin-3-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 410.2; R_(t) = 1.48 ¹HNMR (400 MHz, METHANOL-d₄) δ 8.46 (dd, J = 5.0, 1.7 Hz, 1 H), 7.90 (dd,J = 7.8, 1.7 Hz, 1 H), 7.35 (dd, J = 7.9, 5.1 Hz, 1 H), 4.28-4.20 (m, 1H), 3.85 (d, J = 8.7 Hz, 1 H), 3.71 (d, J = 8.8 Hz, 1 H), 3.51 (s, 3 H),3.50-3.42 (m, 2 H), 3.19-3.10 (m, 1 H), 3.10- 2.99 (m, 2 H), 2.56 (s, 3H), 2.02-1.87 (m, 2 H), 1.81- 1.66 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H).0.096 68

  (S)-6-(4-amino-2-oxa-8- azaspiro[4.5]decan-8-yl)-3-(2-chloro-3-methoxyphenyl)- 5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 445; R_(t) = 0.59 min ¹H NMR(400 MHz, METHANOL-d₄) δ 7.40-7.28 (m, 1 H), 7.17 (dd, J = 8.3, 1.2 Hz,1 H), 7.05 (dd, J = 7.6, 1.3 Hz, 1 H), 4.12 (dd, J = 9.0, 6.5 Hz, 1 H),3.93 (s, 3 H), 3.89- 3.74 (m, 2 H), 3.61-3.42 (m, 6 H), 3.19 (t, J = 5.8Hz, 1 H), 3.12-2.97 (m, 2 H), 2.00- 1.78 (m, 2 H), 1.65 (t, J = 15.1 Hz,2 H). 0.025 69

  (S)-6-(4-amino-2-oxa-8- azaspiro[4.5]decan-8-yl)-5-methyl-3-(pyridin-4-yl)-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one A (M +H)⁺ = 382; R_(t) = 1.20 min ¹H NMR (400 MHz, DMSO- d₆) δ 8.73-8.57 (m, 2H), 8.43- 8.30 (m, 2 H), 3.96 (dd, J = 8.5, 6.6 Hz, 1 H), 3.77-3.54 (m,2 H), 3.47 (s, 3 H), 3.44- 3.35 (m, 2 H), 3.08 (t, J = 6.1 Hz, 1 H),3.03-2.88 (m, 2 H), 1.77 (dt, J = 38.1, 10.2 Hz, 2 H), 1.50 (s, 2 H).0.039 70

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methyl- 3-(pyridin-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 396; R_(t) = 0.60 min ¹H NMR(400 MHz, DMSO- d₆) δ 8.63 (s, 2 H), 8.39-8.31 (m, 2 H), 4.11-4.02 (m, 1H), 3.66 (d, J = 8.5 Hz, 1 H), 3.54- 3.42 (m, 5 H), 3.12-2.94 (m, 2 H),2.91 (d, J = 5.1 Hz, 1 H), 1.94-1.83 (m, 1 H), 1.76 (t, J = 9.4 Hz, 1H), 1.67-1.52 (m, 3 H), 1.09 (d, J = 6.4 Hz, 3 H). 0.008 71

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(dimethylamino)pyridin-4- yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 473; R_(t) = 0.73 min ¹H NMR(400 MHz, DMSO- d₆) δ 8.18 (d, J = 4.9 Hz, 1 H), 6.98 (d, J = 4.9 Hz, 1H), 4.14- 3.99 (m, 1 H), 3.66 (d, J = 8.5 Hz, 1 H), 3.49 (d, J = 8.5 Hz,1 H), 3.38 (s, 3 H), 3.13-2.97 (m, 3 H), 2.94 (s, 6 H), 2.91 (d, J = 5.1Hz, 1 H), 1.96-1.81 (m, 1 H), 1.80-1.70 (m, 1 H), 1.67- 1.49 (m, 3 H),1.08 (d, J = 6.4 Hz, 3 H). 0.022 72

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(ethylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 473; R_(t) = 0.69min ¹H NMR (400 MHz, DMSO- (d₆) δ 8.00 (d, J = 5.0 Hz, 1 H), 6.60 (d, J= 5.0 Hz, 1 H), 6.54 (t, J = 5.6 Hz, 1 H), 4.03-4.09 (m, 1 H), 3.66 (d,J = 8.5 Hz, 1 H), 3.54-3.40 (m, 3 H), 3.37 (s, 3 H), 3.11-2.92 (m, 2 H),2.91 (d, J = 5.1 Hz, 1 H), 1.91-1.69 (m, 2 H), 1.67-1.49 (m, 3 H), 1.17(t, J = 7.1 Hz, 3 H), 1.08 (d, J = 6.4 Hz, 3 H). 0.047 73

  6-((1R,3R)-1-amino-3-methyl-8- azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(dimethylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 471; R_(t) = 0.70min ¹H NMR (400 MHz, METHANOL-d₄) δ 8.14 (d, J = 5.0 Hz, 1 H), 6.99 (d,J = 4.9 Hz, 1 H), 3.50 (s, 6 H), Hz, 1 H), 3.02 (s, 6 H), 2.98-2.83 (m,2 H), 2.32-1.61 (m, 5 H), 1.54-1.12 (m, 4 H), 1.06 (d, J = 6.8 Hz, 3 H).0.028 74

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-((cyclopropylmethyl)amino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one D (M + H)⁺ =499; R_(t) = 0.65 min ¹H NMR (400 MHz, DMSO- d₆) δ 7.99 (d, J = 5.0 Hz,1 H), 6.61 (d, J = 5.0 Hz, 1 H), 6.57 (t, J = 5.7 Hz, 1 H), 4.14-4.03(m, 1 H), 3.65 (d, J = 8.5 Hz, 1 H), 3.51-3.45 (m, 2 H), 3.38 (s, 3 H),3.30-3.26 (m, 2 H), 3.14- 2.78 (m, 3 H), 1.81 (dt, J = 44.3, 9.8 Hz, 3H), 1.67- 1.50 (m, 2 H), 1.18 (ddd, J = 12.7, 7.6, 5.5 Hz, 1 H), 1.09(d, J = 6.4 Hz, 3 H), 0.50-0.38 (m, 2H), 0.33-0.22 (m, 2H). 0.039 75

  6-((3S,4S)-4-amino-3-methyl-2- oxa-8-azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(cyclopropylamino)pyridin-4- yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one D (M + H)⁺ = 485; R_(t) = 0.64 min ¹H NMR(400 MHz, DMSO- d₆) δ 8.07 (d, J = 5.0 Hz, 1 H), 6.68 (d, J = 5.0 Hz, 1H), 6.63 (d, J = 3.0 Hz, 1 H), 4.11-4.00 (m, 1 H), 3.66 (d, J = 8.5 Hz,1 H), 3.49 (d, J = 8.5 Hz, 1 H), 3.37 (s, 3 H), 3.12-2.94 (m, 3 H), 2.91(d, J = 5.1 Hz, 1 H), 2.84-2.72 (m, 1 H), 1.91- 1.81 (m, 1 H), 1.80-1.70(m, 1 H), 1.65-1.55 (m, 3 H), 1.08 (d, J = 6.4 Hz, 3 H), 0.76-0.66 (m, 2H), 0.61-0.52 (m, 2 H). 0.036 76

  3-(2-amino-3-chloropyridin-4-yl)-6- ((1R,3R)-1-amino-3-methyl-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one E (M + H)⁺ = 443; R_(t) = 0.56min ¹H NMR (400 MHz, METHANOL-d₄) δ 7.92 (d, J = 5.1 Hz, 1 H), 6.76 (d,J = 5.2 Hz, 1 H), 3.50 (s, 5 H), 3.14- 2.99 (m, 2 H), 2.96-2.82 (m, 1H), 2.27-1.76 (m, 5 H), 1.55- 1.39 (m, 2 H), 1.30 (dd, J = 12.9, 9.2 Hz,1 H), 1.16 (dt, J = 12.2, 9.9 Hz, 1 H), 1.07 (d, J = 6.5 Hz, 3 H). 0.03277

  6-((1R,3R)-1-amino-3-methyl-8- azaspiro[4.5]decan-8-yl)-3-(3-chloro-2-(methylamino)pyridin-4-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin- 4-one D (M + H)⁺ = 457; R_(t) =0.55 min ¹H NMR (400 MHz, METHANOL-d₄) δ 7.99 (d, J = 5.2 Hz, 1 H), 6.67(d, J = 5.2 Hz, 1 H), 3.49 (s, 5 H), 3.13- 2.86 (m, 6 H), 2.34-1.61 (m,5 H), 1.54-1.36 (m, 2 H), 1.33- 1.22 (m, 1 H), 1.17 (dt, J = 12.3, 9.8Hz, 1 H), 1.08 (d, J = 6.5 Hz, 3 H). 0.030 78

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2-chloro-3-methylpyridin-4-yl)-5-methyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 444; R_(t) = 0.60 min ¹H NMR (400 MHz,METHANOL-d₄) δ 8.25 (d, J = 5.0 Hz, 1 H), 7.46 (d, J = 5.0 Hz, 1 H),4.33-4.17 (m, 1 H), 3.62 (d, J = 8.5 Hz, 1 H), 3.47 (d, J = 8.5 Hz, 1H), 3.58-3.42 (m, 5 H), 3.21-2.94 (m, 3 H), 2.40 (s, 3 H), 2.01-1.85 (m,2 H), 1.79-1.66 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H). 0.017 79

  6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-3-(2,5-dichloropyridin-4-yl)-5-methyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 464; R_(t) = 1.02 min ¹H NMR (400 MHz, METHANOL-d₄) δ8.49 (s, 1 H), 7.70 (s, 1 H), 4.29-4.13 (m, 1 H), 3.61 (d, J = 8.5 Hz, 1H), 3.46 (d, J = 8.5 Hz, 1 H), 3.57-3.41 (m, 5 H), 3.22- 2.96 (m, 3 H),2.00-1.85 (m, 2H), 1.80-1.66 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H). 0.03080

  6-((3S,4S)-4-amino-3-methyl-2- oxa-8-azaspiro[4.5]decan-8-yl)-3-(2-amino-5-chloropyridin-4-yl)-5- methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one A (M + H)⁺ = 445; R_(t) = 0.61 min ¹H NMR (400 MHz,METHANOL-d₄) δ 7.97 (s, 1 H), 6.72 (s, 1 H), 4.20-4.26 (m, 1 H), 3.67(d, J = 8.5 Hz, 1 H), 3.49 (d, J = 8.5 Hz, 1 H), 3.55-3.40 (m, 5 H),3.22- 2.92 (m, 3 H), 2.06-1.86 (m, 2 H), 1.79-1.66 (m, 2 H), 1.22 (d, J= 6.5 Hz, 3 H). 0.129 81

  6-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-3-(2-chloro-3-methoxyphenyl)-5-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one A(M + H)⁺ = 458; R_(t) = 1.40 min ¹H NMR (400 MHz, METHANOL-d₄) δ 7.41-7.28 (m, 1 H), 7.17 (dd, J = 8.3, 1.2 Hz, 1 H), 7.05 (dd, J = 7.6, 1.3Hz, 1 H), 4.23 (p, J = 6.4 Hz, 1 H), 3.93 (s, 3 H), 3.85 (d, J = 8.7 Hz,1 H), 3.71 (d, J = 8.7 Hz, 1 H), 3.54-3.40 (m, 5 H), 3.19-2.98 (m, 3 H),2.03- 1.87 (m, 2 H), 1.80-1.63 (m, 2 H), 1.22 (d, J = 6.5 Hz, 3 H).0.047 82

  3-(2-amino-3-chloropyridin-4-yl)- 6-((1R,3R)-1-amino-3-methyl-8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin- 4-one E (M + H)⁺ = 443; R_(t) =0.56 min ¹H NMR (400 MHz, METHANOL-d₄) δ 7.92 (d, J = 5.1 Hz, 1 H), 6.76(d, J = 5.2 Hz, 1 H), 3.50 (s, 5 H), 3.13- 2.99 (m, 2 H), 2.96-2.86 (m,1 H), 2.28-1.76 (m, 5 H), 1.51- 1.39 (m, 2 H), 1.33-1.27 (m, 1 H),1.19-1.11 (m, 1 H), 1.07 (d, J = 6.5 Hz, 3 H). 0.032 83

  6-(4-(aminomethyl)-4-methylpiperidin-1-yl)-3-((2,3-dichlorophenyl)thio)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one F (M + H)⁺ = 453;R_(t) = 1.03 min ¹H NMR (400 MHz, DMSO- d₆) δ 7.47 (dd, J = 8.0, 1.4 Hz,1 H), 7.21 (t, J = 8.0 Hz, 1 H), 6.94 (dd, J = 8.1, 1.4 Hz, 1 H), 3.33(s, 3 H), 3.28-3.21 (m, 2 H), 3.17-3.00 (m, 2 H), 2.44 (s, 2 H),1.69-1.49 (m, 2 H), 1.43-1.24 (m, 2 H), 0.93 (s, 3 H). 0.008 84

  3-((2-amino-3-chloropyridin-4-yl)thio)-6-((3S,4S)-4-amino-3-methyl-2-oxa- 8-azaspiro[4.5]decan-8-yl)-5-methyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin- 4-one F (M + H)⁺ = 477; R_(t) =0.72 min ¹H NMR (400 MHz, METHANOL-d₄) δ 7.55 (d, J = 5.6 Hz, 1 H), 6.02(d, J = 5.6 Hz, 2 H), 4.37-4.16 (m, 1 H), 3.84 (d, J = 8.7 Hz, 1 H),3.70 (d, J = 8.7 Hz, 1 H), 3.48 (s, 3 H), 3.20-3.06 (m, 2 H), 3.03 (d, J= 5.0 Hz, 2 H), 2.08-1.85 (m, 2 H), 1.83-1.65 (m, 2 H), 1.22 (d, J = 6.5Hz, 3 H). 0.027 85

  (3S,4S)-8-(3-(2,3-dichloropyridin-4-yl)-1H-pyrazolo[3,4-b]pyrazin-6-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine Similar to Example 12 usingappropriate starting materials (M + H)⁺ = 434; R_(t) = 0.98 min ¹H NMR(400 MHz, METHANOL-d₄) δ 8.32- 8.44 (m, 2 H), 7.84-7.75 (m, 1 H),4.31-4.09 (m, 3 H), 3.93- 3.85 (m, 1 H), 3.77-3.70 (m, 1 H), 3.57-3.39(m, 3 H), 3.05- 2.98 (m, 1 H), 1.95-1.61 (m, 5 H), 1.22 (d, J = 6.6 Hz,3 H) 0.067

Assays

Compounds of the invention were assessed for their ability toselectively inhibit SHP2 activity. The inhibitory properties of thecompounds of the invention described herein can be evidenced by testingin any one of the following assays.

SHP2 Allosteric Inhibition Assay

SHP2 is allosterically activated through binding ofbis-tyrosyl-phosphorylated peptides to its Src Homology 2 (SH2) domains.The latter activation step leads to the release of the auto-inhibitoryinterface of SHP2, which in turn renders the SHP2 protein tyrosinephosphatase (PTP) active and available for substrate recognition andreaction catalysis. The catalytic activity of SHP2 was monitored usingthe surrogate substrate DiFMUP in a prompt fluorescence assay format.

More specifically, the phosphatase reactions were performed at roomtemperature in 384-well black polystyrene plate, flat bottom, lowflange, non-binding surface (Corning, Cat#3575) using a final reactionvolume of 25 μL and the following assay buffer conditions: 60 mM HEPES,pH 7.2, 75 mM NaCl, 75 mM KCl, 1 mM EDTA, 0.05% P-20, 5 mM DTT.

The inhibition of SHP2 by compounds of the invention (concentrationsvarying from 0.003-100 μM) was monitored using an assay in which 0.5 nMof SHP2 was incubated with of 0.5 μM of peptide IRS1_pY1172(dPEG8)pY1222(sequence: H2N-LN(pY)IDLDLV(dPEG8)LST(pY)ASINFQK-amide) (SEQ ID NO:1).After 30-60 minutes incubation at 25° C., the surrogate substrate DiFMUP(Invitrogen, cat# D6567) was added to the reaction and incubated at 25°C. for 30 minutes. The reaction was then quenched by the addition of 5μl of a 160 μM solution of bpV(Phen) (Enzo Life Sciences cat#ALX-270-204). The fluorescence signal was monitored using a microplatereader (Envision, Perki-Elmer) using excitation and emission wavelengthsof 340 nm and 450 nm, respectively. The inhibitor dose response curveswere analyzed using normalized IC₅₀ regression curve fitting withcontrol based normalization. IC₅₀ results for compounds of the inventionare shown in examples and tables 1-7, above.

p-ERK Cellular Assay

p-ERK cellular assay using the AlphaScreen® SureFire™ Phospho-ERK 1/2Kit (PerkinElmer): KYSE-520 cells (30,000 cells/well) were grown in96-well plate culture overnight and treated with Shp2 inhibitors atconcentrations of 20, 6.6, 2.2, 0.74, 0.24, 0.08, 0.027 μM for 2 hrs at37° C. Incubations were terminated by addition of 30 μL of lysis buffer(PerkinElmer) supplied with the SureFire phospho-extracellularsignal-regulated kinase (pERK) assay kit (PerkinElmer). Samples wereprocessed according to the manufacturer's directions. The fluorescencesignal from pERK was measured in duplicate using a 2101 multilabelreader (Perkin Elmer Envision). The percentage of inhibition wasnormalized by the total ERK signal and compared with the DMSO vehiclecontrol.

Colony Formation Assay and Cell Proliferation Assay

KYSE-520 Cells (1500 cells/well) were plated onto 24-well plates in 300μL medium (RPMI-1640 containing 10% FBS, Lonza). For drug treatment,compounds of the invention at various concentrations (20, 10, 5, 2.5,1.25 μM) were added 24 hours and 5 days after cell plating. At day 11,colonies were stained with 0.2% crystal violet (MP Biomedicals) andsubsequently dissolved in 20% acetic acid for quantitation using aSpectramax reader (Thermo Scientific). In cell proliferation assay,cells (1500-cells/well) were plated onto 96-well plates in 100 μL medium(RPMI-1640 containing 10% FBS, Lonza). At day 6, 50 μL Celltiter-Gloreagent (Promega) was added, and the luminescent signal was determinedaccording to the supplier's instruction (Promega).

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims.

1. A compound of formula I:

in which: Y₁ is selected from N, NH and CH; Y₂ is selected from N, NHand CH; Y₃ is selected from N and C; Y₄ is selected from N and CH; R₁ isselected from R_(1a) and SR_(1a); wherein R_(1a) is selected fromphenyl, pyridinyl, pyrazine, pyridazine, 2,3-dihydrobenzofuran andpyrimidine; wherein said phenyl, pyridinyl, pyrazine, pyridazine orpyrimidine is unsubstituted or substituted with 1 to 3 R₄ groups;wherein each R₄ group is independently selected from halo, amino,—NH(C₁₋₄ alkyl), —N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl),morpholino, pyrrolidinyl, hydroxyl, C₁₋₃ alkyl, C₁₋₃alkoxy andhalo-substituted-C₁₋₃ alkyl; wherein said pyrrolidinyl of R₄ can beunsubstituted or substituted with hydroxy; R₂ is selected from H, CH₃and halo-substituted-C₁₋₂ alkyl; R_(4a) and R_(4b) are eachindependently selected from hydrogen, hydroxy and fluoro; with provisothat R_(4a) and R_(4b) cannot both be OH; R_(5a) is selected from aminoand amino-methyl; R_(5b) is selected from C₁₋₆ alkyl, methoxy-carbonyl,C₃₋₆ cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂ alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl, halo-substituted-C₁₋₃ alkyl, halo-substituted-C₁₋₃ alkoxy andC₁₋₃ alkoxy; and R₁₃ is selected from hydrogen, halo and C₁₋₂ alkyl; andR_(6a) and R_(6b) are each independently selected from hydrogen, hydroxyand fluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; orthe pharmaceutically acceptable salts thereof.
 2. The compound of claim1 selected from Formulae Ia, Ib, Ic, Id, Ie, If and Ig:

in which: R₁ is R_(1a), wherein R_(1a) is selected from phenyl,2,3-dihydrobenzofuran and pyridinyl; wherein said phenyl or pyridinyl isunsubstituted or substituted with 1 to 3 R₄ groups; wherein each R₄group is independently selected from halo, amino, —NH(C₁₋₄ alkyl),—N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl), morpholino,pyrrolidinyl, C₁₋₃ alkyl, C₁₋₃ alkoxy and halo-substituted-C₁₋₃alkyl;wherein said pyrrolidinyl of R₄ can be unsubstituted or substituted withhydroxy; R₂ is selected from H and CH₃; R_(4a) and R_(4b) are eachindependently selected from hydrogen, hydroxy and fluoro; with provisothat R_(4a) and R_(4b) cannot both be OH; R_(5a) is selected from aminoand amino-methyl; R_(5b) is selected from C₁₋₆ alkyl, methoxy-carbonyl,C₃₋₆ cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂ alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂ alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; or thepharmaceutically acceptable salts thereof.
 3. The compound of claim 2 offormula Ia:

in which: R₁ is phenyl substituted with 1 to 3 R₄ groups; wherein eachR₄ group is independently selected from halo, amino, —NH(C₁₋₄alkyl),—N(C₁₋₄alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅ cycloalkyl), morpholino,pyrrolidinyl, C₁₋₃ alkyl, C₁₋₃ alkoxy and halo-substituted-C₁₋₃ alkyl;wherein said pyrrolidinyl of R₄ can be unsubstituted or substituted withhydroxy; R₂ is selected from H and CH₃; R_(4a) and R_(4b) are eachindependently selected from hydrogen, hydroxy and fluoro; with provisothat R_(4a) and R_(4b) cannot both be OH; R_(5a) is selected from aminoand amino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃ alkyl, hydroxy-substituted C₁₋₃alkyl,C₁₋₂alkoxy-substituted C₁₋₃ alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂ alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl, halo-substituted-C₁₋₃ alkyl, halo-substituted-C₁₋₃ alkoxy andC₁₋₃alkoxy; and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; andR_(6a) and R_(6b) are each independently selected from hydrogen, hydroxyand fluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; orthe pharmaceutically acceptable salts thereof.
 4. The compound of claim3, or a pharmaceutically acceptable salt thereof, selected from:


5. The compound of claim 2 of formula Ic: in which:

R₁ is selected from phenyl, 2,3-dihydrobenzofuran and pyridinyl; whereinsaid phenyl or pyridinyl is unsubstituted or substituted with 1 to 3 R₄groups; wherein each R₄ group is independently selected from halo,amino, —NH(C₁₋₄ alkyl), —N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl),morpholino, pyrrolidinyl, C₁₋₃ alkyl, C₁₋₃alkoxy andhalo-substituted-C₁₋₃ alkyl; wherein said pyrrolidinyl of R₄ can beunsubstituted or substituted with hydroxy; R₂ is selected from H andCH₃; R_(4a) and R_(4b) are each independently selected from hydrogen,hydroxy and fluoro; with proviso that R_(4a) and R_(4b) cannot both beOH; R_(5a) is selected from amino and amino-methyl; R_(5b) is selectedfrom C₁₋₆alkyl, methoxy-carbonyl, C₃₋₆cycloalkyl-C₁₋₃alkyl,hydroxy-substituted C₁₋₃alkyl, C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5to 6 member heteroaryl ring containing 1 to 4 heteroatoms selected fromO, S and N; wherein said C₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃alkylof R_(5b) is unsubstituted or substituted with 1-3 fluorines; or R_(5a)and R_(5b), together with the carbon atom to which R_(5a) and R_(5b) areattached, form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂ alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; or thepharmaceutically acceptable salts thereof.
 6. The compound of claim 5,or a pharmaceutically acceptable salt thereof, selected from:


7. The compound of claim 2 of formula Id:

in which: R₁ is selected from phenyl, 2,3-dihydrobenzofuran andpyridinyl; wherein said phenyl or pyridinyl is unsubstituted orsubstituted with 1 to 3 R₄ groups; wherein each R₄ group isindependently selected from halo, amino, —NH(C₁₋₄ alkyl),—N(C₁₋₄alkyl)₂, —NH(C₀₋₁alkyl-C₃-5cycloalkyl), morpholino, pyrrolidinyl,C₁₋₃alkyl, C₁₋₃alkoxy and halo-substituted-C₁₋₃alkyl; wherein saidpyrrolidinyl of R₄ can be unsubstituted or substituted with hydroxy; R₂is selected from H and CH₃; R_(4a) and R_(4b) are each independentlyselected from hydrogen, hydroxy and fluoro; with proviso that R_(4a) andR_(4b) cannot both be OH; R_(5a) is selected from amino andamino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆cycloalkyl-C₁₋₃alkyl, hydroxy-substituted C₁₋₃ alkyl, C₁₋₂alkoxy-substituted C₁₋₃alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆ alkyl or C₁₋₂ alkoxy-substituted C₁₋₃alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂ alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH; or thepharmaceutically acceptable salts thereof.
 8. The compound of claim 7,or a pharmaceutically acceptable salt thereof, selected from:


9. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein the compound of claim 1 is:


10. A compound, or a pharmaceutically acceptable salt thereof, selectedfrom formula II:

in which: Y₅ is selected from N and CH; Y₆ is selected from CH₂ and NH;R₁ is selected from R_(1a) and −SR_(1a); wherein R_(1a) is selected fromphenyl, pyridinyl, pyrazine, pyridazine, 2,3-dihydrobenzofuran andpyrimidine; wherein said phenyl, pyridinyl, pyrazine, pyridazine orpyrimidine is unsubstituted or substituted with 1 to 3 R₄ groups;wherein each R₄ group is independently selected from halo, amino,—NH(C₁₋₄ alkyl), —N(C₁₋₄ alkyl)₂, —NH(C₀₋₁alkyl-C₃₋₅cycloalkyl),morpholino, pyrrolidinyl, hydroxyl, C₁₋₃alkyl, C₁₋₃alkoxy andhalo-substituted-C₁₋₃ alkyl; wherein said pyrrolidinyl of R₄ can beunsubstituted or substituted with hydroxy; R_(4a) and R_(4b) are eachindependently selected from hydrogen, hydroxy and fluoro; with provisothat R_(4a) and R_(4b) cannot both be OH; R_(5a) is selected from aminoand amino-methyl; R_(5b) is selected from C₁₋₆alkyl, methoxy-carbonyl,C₃₋₆ cycloalkyl-C₁₋₃ alkyl, hydroxy-substituted C₁₋₃ alkyl,C₁₋₂alkoxy-substituted C₁₋₃ alkyl and a 5 to 6 member heteroaryl ringcontaining 1 to 4 heteroatoms selected from O, S and N; wherein saidC₁₋₆alkyl or C₁₋₂alkoxy-substituted C₁₋₃ alkyl of R_(5b) isunsubstituted or substituted with 1-3 fluorines; or R_(5a) and R_(5b),together with the carbon atom to which R_(5a) and R_(5b) are attached,form a group selected from:

wherein *C represents the carbon atom to which R_(5a) and R_(5b) areattached; R₁₀ is amino; R_(11a) is selected from hydrogen, hydroxy,fluoro, C₁₋₂alkyl and hydroxy-methyl; R_(11b) is selected from fluoro,methyl and hydrogen; R_(11c) is selected from hydrogen, C₁₋₂ alkyl andhydroxy-methyl; R₁₂ is selected from hydrogen, halo, hydroxy, C₁₋₃alkyl,halo-substituted-C₁₋₃alkyl, halo-substituted-C₁₋₃alkoxy and C₁₋₃alkoxy;and R₁₃ is selected from hydrogen, halo and C₁₋₂alkyl; and R_(6a) andR_(6b) are each independently selected from hydrogen, hydroxy andfluoro; with proviso that R_(6a) and R_(6b) cannot both be OH.
 11. Thecompound of claim 10, or a pharmaceutically acceptable salt thereof,selected from:


12. A pharmaceutical composition comprising a compound of claim 1 or apharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier.
 13. A method of treatmentcomprising administering a compound of claim 1, or a pharmaceuticallyacceptable salt thereof, to a person in need of such treatment in aneffective amount for the prophylactic or therapeutic treatment of adisease or disorder which is mediated by the activity of SHP2.
 14. Themethod of claim 13, wherein the disease or disorder mediated by theactivity of SHP2 is selected from Noonan Syndrome, Leopard Syndrome,juvenile myelomonocytic leukemias, neuroblastoma, melanoma, acutemyeloid leukemia, breast cancer, esophageal cancer, lung cancer, coloncancer, head cancer, neuroblastoma, squamous-cell carcinoma of the headand neck, gastric carcinoma, anaplastic large-cell lymphoma andglioblastoma.